Oscar Leonardo Mosquera Dussan. Ingeniero Electrnico. Universidad de la Sabana. Facultad de Ingeniera. Doctorado en Biociencias.

Componente gentico de Epilepsia en lbulo temporal

Introduccin Estudios recientes reportan mutacin de la protena leucine-rich glioma-inactivated-1 (LGI1), como causa de epilepsia parcial en humanos [1-3], anormalidad en el gen LGI1 asociado a la protena es responsable de epilepsia autonmica dominante de lbulo temporal [4-6]. Tambin se presenta un rol del gen LGI1 en epilepsias autoinmunes [7]. En la actualidad, poco se sabe sobre el patrn de expresin temporal y espacial del gen LGI1 durante la embriognesis normal, un reciente estudio con un ratn transgnico implican que Lgi1 puede tener un papel en el establecimiento de la arquitectura normal del cerebro y funciones neuronales durante el desarrollo cerebral lo que sugiere que pueden estar implicados en la neurognesis y la plasticidad neuronal, que se define ms especficamente en el animal adulto [8]. Otro estudio en animales muestra que el truncamiento del gene LGI2 (homologo al gen LGI1 en humanos) muestra una remisin de epilepsia focal en perros [9]; estudios sugieren que ambos genes presentan una gran similitud en los mecanismos moleculares de la patognesis [10]. En el presente trabajo se realiza un estudio bioinformtico al gen LGI1, se estudian las secuencias de ADN, ARNm, y la secuencia de aminocidos que forman la protena leucinerich glioma-inactivated-1.

En ncbi se puede apreciar la informacin del gen LGI1 asociado a epilepsia de lbulo temporal.

EL gen se encuentra ubicado en el cromosoma 10 y la presente secuencia no tiene splicing alternativo.

La secuencia del gen en formato FASTA es la siguiente,

>gi|224589801:95517566-95557916 Homo sapiens chromosome 10, GRCh37.p5 Primary Assembly GAAAGCAGAAAGCTGTTTATTTTCTGTGACATCACTTCACTTTGCCTTCGAAGGCTGGTCTGTGCTCAGT GTTTTCGTGGTGATGCAAGTCGGCTCTCTCCTCCAGCAGTTGGATCCCTCCCATCTCACAGTACCTCACA GGTCTCTTCCCCCGAGCAGTGCATTGCTGGAGCGAGGAGAAGCTCACGAATCAGCTGCAGGTCTCTGTTT TGAAAAAGCAGAGATACAGAGGCAGAGGAAAAGGGTGGACTCCTATGTGACCTGTTCTTAGAGCAAGACA ATCACCATCTGAATTCCAGAAGCCCTGTTCATGGTTGGGGATATTTTCTCGACTGCATGGAATCAGAAAG AAGCAAAAGGATGGGAAATGCCTGCATTCCCCTGAAAAGAATTGCTTATTTCCTATGTCTCTTATCTGCG CTTTTGCTGACTGAGGGGAAGAAACCAGCGAAGCCAAAATGCCCTGCCGTGTGTACTTGTACCAAAGATA ATGCTTTATGTGAGAATGCCAGATCCATTCCACGCACCGTTCCTCCTGATGTTATCTCATTGTAAGGCCC GTAAGCATTTTGATATCTAATTTACGATTTAAAAATTCCAGCCGGTGGATTTGGGGCTTTGCATGTATTT GTAGAAGGGCATGGAGAGAGAGATTCCTCTTGCATGCTTGGCCATTTGACAGTGCTAACATTTGCTGCAT TTAGAATTTTTGATTTTTTTAGCTGCTACTGAACATGCTTAGATACCCCCAGCCCTGAACATTATCATGA GAAACCTGTAGCCGATTCATTTCTCTTACTTCATCTGGGAAGGAATAGTATCGTACTTCATAAAATAGTG TCAAAATAGTCACTGTTATGCTAAACCGGATTAACATAAGGTTTGTTCTGTGTGTGTGTGTCTCTTTGTG TGTGTCTGTTTGTTTGTTTTCTCTTTTTTGTTTTCTTTCAGATCCTTTGTGAGATCTGGTTTTACTGAAA TCTCAGAAGGGAGTTTTTTATTCACGCCATCGCTGCAGCTCTTGTGAGAAATATTTATATCATGACTATT

TTTAATATGGCATATATTTGGATAAGCCTTCTAGTAAAATGATCTCAATATTAATTTTGTCAAATGTGAT TCTATTTCTGAGAAAACAGAATATCCGTAAAGTGAGAGGTAGATGGGTTTGTTTGCGACTTCACACCAAC AGTGCAGGTTGATTCTTACAATGGTAAATCTGTTAACCTGTCATGCTGTCTACTTTAATGGCTATTTATG AAGTTGTTGGAATCTTGCAACAATGATCCCAATTCCATTCCACTATTTTACTTTCATTTTTAACTTGGGA ACTTGGGTTCTTTTGCCCTGGAGTTTTTGAATTAGTCAATACAGAACCCAGCATAACATATGCGCTCAAG ATGTTGTCTCCGGGGATCTGGGGCAGGCTGCTACTCTGTATGATTTACCATGTGGGGAAAAAAAACACCT CAGCTCTTTCTAGGTAATTCCTTAAAATGTTAATTCTCAAGAGACTGTTTTTTAATCTCATCTAATTTAT AACTAGTTCCATTACAAGGAGGGAAAGAAAATATTAATATAGGATAGTGCAAAGGATTACTCTCTGCTAT ATGTTTTGAGTTGGCCACAAGGGAACCACATACTCAGTCTATGCCTGAAAATATTCAAAAAAATAATATT TGACTAAGAAATCAGATCTGTTCACAGGCATAATGGAACACATAAAATTTGTTCTGATGTGTAATGTTTG GCAATTGGAGTCCTAGTCCTTACAGCGGTATCACATGGGGGTCAAGAGGTCTCCCAGGACTCAGGTATCA GGTCGCAGACTTGGTTTGTAGCTGGACGGATATGGGCAGAATTAGACAATTGTGGATCAGCTGAGTCTTG GCTATCTTAACTGCATGATTCAGGACCCAGGCCTTTCACTTGTTTGTTTTTTATTTTTTTATTTTCTTGC CTTACTTGGCCATTTTGGCTGTTTGAAGTTACATACACAGAGGCAAACAGCATGGAGTGGACGTTGGAGA GACAAGTGTTTAAACCTCACCCTGCCACTCATAGAACCTAAACATGTAGTCTTGTGCAGATTAGTATATC TCACTAAGCCTCAGTCTTCTTATCTTTAAAATGGAAGTGAGAACACATACTTCCAAGGTTTAAGTAGATA AAACACTTGGCATAGGGTGTGCCAAATGCTTACACATTTATAAATGGTAGGGTTTAAAAATTTTATATCA AGCTAAAAAGCTAAAAGTCACTCATCTATTGTTTTGTGTATATGAAATTCCTAAACTGGTTTGACTGTTT CATTGGATTCACAAACTTGATCAGTTGTTTTGACATTATGTAATGATGTTGGCTAATGCCCTTCTACTGG CTGATGTCTTCAAATGATTAGCAATTGACTAGATATTCCTAAGGGGTTTATCTGTTTGATTGATGGCTAG TGGTGTGACTGGGGGAAGTGTTAGTTAGCACATCAAACAAATCCAAAAAAGATAGATTCTTTCCCACAGA AAGGGGCAGAGTCTCTAATGCTGGGATCCTTCCATAGGATAAGAATTTGCTGTCTCTGAGAAAGTGATAA ATTGTGTTTCTGGTCCTCACTTGTTAAATGAATAAATAGCTCTTTTAAAAAGCATCCTTTTGATATGCTA AGGCACCTCCCTTCCCCTAGGAAGTTACATAGATCTAGTTACATAGATCAGTCCTTAAAATTCCAGGAGT AGTGTCAGTAGACACTAGATGGACATTAGCTGTGGCACAGAAAGCCTGGAGTGATTAACCTAATCAATTA CCACATCTCCACTTTGTTTTTGTTGGGTGGACCCAGTCACGGCAAGCACAGATGCTGGACAATGTTATAG AAATAGTTACCGGCTGCTGCAGGGTAATGTGATTTCACGTATTTAGAATCACAGAGGGAAGCCAAGGGAC TGGTTTTTAAATAGCCCAACCGAGTGTGTGCCATGCCCGTGCCCCATGCTCCGAGTTCCTATGGAACATT CCAGCCTTTATTTCTAAAAGCAGTTCAGGTTTCAGAAATAACAAAAGGCGAGTAATTCTCTCCCAGTGTG TTCTCGGTTAACTCAGATTGGCTAACCTCAACATGGTAGACTTGTGCCAGAGCTGAACTTATGCAAAATT GTCAGGGGCCAAAGTTTGTCCCTAATCTCCAGGACCCTGTGCCTGAAGACTCTGCTCTCCATCCTTCTTG CCCAATATTCTCTGCTTCCGCCTATCCTCAGGAAAAGGTCTAATAAACTCATGTGCGGTGTGTAGGAAAT ATATGAGAAGGCTCTGAATATGAACTATGAAACGATCATAACAATTGTGGGAATGCATGGATCCCACAGA GAAAGATCTCTTCTGTGAGACAGGGCTGTCCAGTGCCCTGAAAACCCCATATGGGCTGCCAATCCCCCTT TGGAACATGAAGACCAATGGCATTGGACCACCAGATTAGGCCTAGATCACACAAGCCATTGAGGTTTGCA ATGAATTCCAACAGCATTTGTGATTGTGAATTGCCACACTCACCCATCATACACTGTGAAGAGATAAATT TCAGAATCAATTCGGCTTGATTGACGTAGGAAGACAGTTCGTTGGTTTCTTGCTTTATTGTTGGGAAGGG TTAAAGCAAATGTCCTATTTCCATCATCCACCCTAAAAATCTGATAATGTCAGTATGCCCCCAGATGCTT TTTCAGAGCTTCTGAAAAAGCTATGATGAGAAGTACTTGTTGAAATGTCAGTCTGGGGTACTGAGGAAAA TAGACATGGTATCTGAAGTTACATGCAGTGTTTGGGAAAAGATCCTGGTAGGCCATGGCTGGCAGTTTGT GTTGGAGAACAAGAAAAACAAAAACCCACATTCCGTAGAGAAGAGAGAGAAAGAATAAAATTATTCAATA AAAATTCATTAGATTAGTTGTAATATACATTTTTTAAAAATCTCATTTATGGGTTTAAACATAACAATAA AAACCTTTCCCAAGCAAATGCAATTTCATTCATGTTTTACATAGGGCCAGTTTCGAAACATAACCAATGA CAACGATTATGCGTTCTAGCTTGCTTTTAGTGATAGAAAGTGGTACTTTTAGTGAATGCAGTTTAGGGAA ATGTGTTTTCACACATTACAGCCCTGTTCCTTCTCAACTACAGACTGAAGTCCCCAATGATATGGGCTCA TGATGGAAGTGAAATTCAAATTTAAGAGGACAAAATGTGGCCACTTCGCCACCCACTTATAACATATTTT AATTATAAACCCCTGTAATTCCCGAGAGTGTTCTATTTTCAGGGAGTGATTTCCAGGGTAGTTTTAGTAT CATGCTCAAATAAAACAACACAAAACAAAAACACTGCACGTAAGAATCCGACATGACCAGAAAAAATTTC TTAGTATTCCAATTGGATGTGGTCTTGTTTGAAAGACAGAATTGATCTTTCAGGGTCAAAGGAGGGGAAT AGCGTTATTTTAAAAATTAATCCACAATCAACACATAAATACCTGATCCATAAGAAATTAAAAACTAAAA ACAAATGGAAAAGCAAAAGGCATGAACAAAAGAAGGAATAAGTCCATCCTTACTAAGGCTTTATCATTTA GTCCTTCAAAATATGATCTGTCCAGGAAAAAATATCCATGGCCTCCTTTACCTTTAAAGCAATTGCTACC TTTTTCTATTCCTCCTTATATTAGAAGCCTACAATTTAGCAGTCTTCAATGACCACTCAAGCCAAACCTA TCAGCCAAGGGTGTTTTTGGAGGAATTATTAGCTGGTTATAGGTGAGCACGTTTTGGTTTGTATCCAGCT GATGCCCCCATGATGGACCAAAAAGCAAAAATTTCAGTCATCTAGGCTGTTTTGTCTACAGTTTGCCCAA TGATCCATAGATCTAATCTTTAACTTTTTAATTAATTACTTCTTTTTTGTTTTTACTTTCTTAAAAATAA TTTTAAATTTTATTTTAGATTCGAGGGTCCATGTACAGGTTTGTTACTTGAAGTGAATTAATTCTCGTAA CTGCACAAACTAGCACTCACCACAATTGTTTTCTCTGAAAGTGTATTTCATCCTTTCAGTTATTCAGGCA AGAAACTTCAGAGTTACCTGACCCCTCTCTTTCACTCTCCACTTCCACCTGGACAAGCATCCTGTCCGAC AGCATGAGAATCTGCCTCCTCCTCTTCACCACAGCACCACTGCTTTCACTGAGGCTGCATTTCATTCACT GTTTCACAGGCTGTGGCAGGAGCCTCCACGCCTCACTGTCTGCTTTCAGTTCCCATCTGACCCATCTCCA TTAGCGCTGCCTTTCCTGAGACACAGCTCTCCGTGCTTTCATGACTCCCTAGAAACAGAGAAAGTCCAGA CACCTGAGCTGGCATCCATAGTGTTTGGCCCTTTCTATTTGTCCAAGGTTATCACTCACATCATTGACTC TACACACACAGACACACACACACACACAGACACACACACACACACGTGCCCAAACAAGTTCTGCAGGATC TCATTCTATGACCTCTGTTGATTCTCAAATTCTCTAGACTTTACTGATGTCTGGCACTGTGTGCCATTTG

CTAGAGTTATGGTGCTTGTATCTGCCTTGTGTCTTTCATGGAGGATCACCAATAAACTGTGGTCCCAGAG AAAAACCCAGACATGTTTTCCCATGTATTTGTTGAATCCAGTTTATGTTTCCATCGATTTTGTGCTGAAG ACTCCTGTGTTACTCTCAGCAGATAATGCTGGTCATGTTCACACTCTTTCTGATGTTTGGTGCACCCTAT CTGAATTCACGTGGACTAATAGTGTGGAGGTCTGGGAAGTGAGCCGAACTAAAAAAACTCGAGGAGATAT TTGCTCATGAAGTTGTGAATTCAAACTTCCTCAGCAGAGATTTCCAGCATACCAGATTTCAAAGGCAAAG CATCTAAGCCTTTCCTTAGACCCACCGCTCGAATCTCTTGTGTCCCACGCTGACTTCACATTTGGTCTCC TTGGTTTTTAGTCAGACTGCAGAGGCCGACTAGTCTGTCCTGGGAAGGGTAAGATGCTCCTTTATCCAGC CTGACCGGACATCCAGAGGGAAATGTTAGTTAAGCATTATTAATATGAAAAGAAAAATCAATAACACAAA GATAATTTCCTTTTTGATTTAAGTACTGCTTAGCCAAAATAAGTATTTATTTATTGTATAGTTGCAAGCA GCAACAAAATGAAAACAAAAGCCTCCCATTTTAGCAGACATGTTTTGAGGCTCTACTGGGTAGCAGGTAC AATGTTGGGCTTTGTGAAGGACACAAAGATGAGTGACACGCTGTCACTTTTCACCCTGATATGGCAGCTC CAAGCTGGTAGTGATTCTTGATATGGTAAATTATACTTTGCCTGTATTTGAATGCATTAGTTTCATTTAA AAAATTATTTTTATGTAAGTAGAGATGAGGTCTTGCTATGTTGCCCACGCTGGTCTCGAACTCCTGGCCT CAAACAATCCTCCCACCTCACCCTTCCAAAGTGCTGGGATTATGGGTATGTGCCTGGCCTCCTTAAATGT ATTATAAGGTAGTCTTGATAAGCATTCTCATTTTGTTCAAAATGAGATTCAGTTGATTAGTGATGATGGT GATGCTTGCTGGTTGTTGATCCTGTGGCTAATACGGGCTACAGAATGTCTGCAAGGGCTTTACAGCATCA GCTGTAATATCAAAACAGCTGACCCTGGTGAGCTGGCAATGAGGCAGACAACACAGACTTGTGGGCAGAT AAGCATGGGTTCAAATCCCCACTCTGCTTCTTCCTAGTTGTATGATTTGGGGCAAGCTTAATCAAGTCTT GATATCTATATCTACAAAATGAGGAAGAACCGTAGTACCTATTGCACAGAATTACAGTAAGGATTAAGTA AAACAATGCATGAAAAATGCTTTGCAAGGTTCCTGTAACTCAATACACATTAGCTTTATTACAGTATCTG AAAATCACTTTAATTGGAACTACCATAACATCAGATTAGTGTCTGATTGTATAATCTGGGTTTGCCGATA ATCTCAGCTTTAAACATTTTGTGGTTCTACACACAGTTCCCCAGCAGAACTTGCTGAGAAATCTTTTCTT TAAATACTTCTTACAGGAAACATGAACTATTTACCTTTCAAAAAAAAGTCATAACCCTCTCCTTTCTTCC TCATGAATAAAATGACTGGCACCACGTTTTGTAAAACATGCCAGATTTTGTTTAAATAATTAATATTGGC AGTCTTCAGATGAGTGGTTCTATGGTAGCATAGAAAGAGGCCTAAACTAGAAATCAGGAAACATGGGTTT GAAACTCAGTTTGGCCACTATTGGGCTGCGTGATCCAATAGTTTGGGTAGGTCACAAAACTACCTAATAG TTTGGGTAGGTCACAAAATATCACTGGACCTCAGCTTTTATTGCATAAAATATTAGAGGTAGATTAAACT GTGGTTCTGAAACTCAAATGCTAGTGGGGTTTGGGGCGGTAAGACCAATGAAGGAAGCAGTTGAGTATAA GACAATAAGGAGTAGTGGTGACTGAGGTAAACCAGAGAAGACATGTCTTACCTAAAAATCCTCTGCTCCA ACTGACTATTGCTGATGGGATTGTGGTACATGCACATGGAGCCCTATCTTCCTCATTTTTAGGGGAAGGT GGAATTCTGGAAGTAGATTTTTAAACAGGATAAAGACCAAACAAATATAAGTTGGCATTTTGAGACCCTT GCATTCAATTACATCTAAGGTCTCCTCCTATCTTAAAATTCTGTTGTATTCTACAAACAGTTACGTTAAT ATAATATAGAAATGGCATGGTTAAAGAAATGGACAACGATGGCTTGGCGCGGTGGCTCACGCCTGTAATC CCAGCACTTTGGGAGGCCGAGGAGGGTAGATCACGAGGTCAGGAGATGGAGAACATCCTGGCTAATACGG TGAAACCCCGTCTCTACTAAAAATACACAAAATTAGCCATGCGTGGTGGCGGGCGCCTGTAGTCCCAGCT ACTCGGGAGGCTGAGGCAGGAGAATGGCGTGAACCCAGGAGGCGGAGCTTGCAGTGAGCCGAGATCGCGC CACTGCACTCCAGCCTGGGCGACAGAGTAAGCCTCCGTCTCAAAAAAAAAAAAAAAAAAAAGAAATGGAC AAAGATTATTGAATCAGTTCTAACAAACCAATACATTAGGTTGTTCATTGAAAGAAACTCATAATATGAA ACTGGCTGTTATAAGGTTATTAATACAATAATTGTAGTTCCCAATCTCCCAAACAGGTCCAAAGGGACCT TACCAGGCTCTCCCTGTGGTGGTTGATGATAAAATGATGTTTCAGTATCCCTCTCATGGAGCAAGGATTG GAAAAAAATTGTTAGGCCCATTTTTATATAGCAAGATGGCAGGTGCTTGCATGTGTAAAATGTGATCAAA AGGCAATAAATCTTTAAAAATACAAACTAGAATGTATACATACAAATAACTCTTGACTTCACAGAGGGCC ATAGATTTTGCCCAACTGTGCTTCCATGGCTAAAGATTTCTGGACCCTCCTTTGAGAATTGTCTTCATTG CCAGTTTACAAGCCACGCAATAAAACCAGGCAAGGTTTTATTAGAGACATTAGTTATTTTTGTATTGCTT GGTGCTAAGCATTTTATAGATCTCTTTTTTTTTTGAGACGGAGTCTCGCTCTGTCCCCCAGGCTGGAGTG CAGTGGCGCGATCTCGGCTCACTGCAAGCTCCGCCTCCCGGGTTCACGCCATTCTCCTGCCTCAGCCTCC CGAGTAGCTGGGACTACAGGCGCCCGCCACCACGCCCGGCTAATTTTTTTGTATTTTTTTAGTAGAGACG GGGTTTCACCGTGTTAGCCAGGATGGTCTCGATTTACTGACCTCGTGATCCTCCCGCCTCGGCCTCCCGA AGTGCTGGGATTACAGGCGTGAGCCACCACGCCCGGCCTATAGATCTCATTTTTAATGTCATCATCATGG TAACCTTTTGAGGTAGATATAATTATCTCCATTTTGCAAATGAAGCGCTGAGTCCTAAAAGATAACCTCC TCAATGTCACGCCCCTATTAAGTAGCAGAGTTGGAGTTTAAACCCATGTCCTCACTTTTAACCACTTCTG TCTACTTCCTCCTGAACATGAATCATCATGCTTCACTTTGCATAGTTTTGTTCCCAAAATCAAATCCCCT CTCAAAGGGTCAACCTTTGCCATTAACAAGAGTTTTCAGGAGAATGTGCCCTAATTCCTGGAGACAATGA CTGAAAGGAGTTTCAAAAGCCCTTTGAAAGAGTGGCCGCATTGCTGGCAAGAGCAGCTAGTTTCCCTAGG TTGCAACTCTCATTTATATGTGTAAGCTCTGATCTCCTTGCTTAAACAAAAATAGAGAGAAAGAAAAGAA AAAAGCAAAAGAAAAAGGCCTTATTTACTTTAAAGTTACACCTTAGACTACTGTTGTCTAAACTTGTGTC CTGTGGAGCAGTTGACCTGAAAGAAGAGTTCTGTGCTCAAATAAATTTGGGAAACTGCACGTTCTATCCC TGCTTGGTTATTTACAATGAAGTTACCTTGGCATAGAAACATCTCTGAAATTATTCAAAATTGTTTCACT TTTCTTAACATCCCATAGAATATTGGTTCTATGGAACATACAGTACTTTGGAAATATATGCATGTGATAA TTCATCTTATTTTTAGCTTAACTATTTGTACCCCACCTCATTCTAGAGAGGTTGGATAGCAAAGGATACG CAGGTGGAAAGAATCTCATCCTAGTGGATACATTAAACGGTCTGTCTCCATATAAGGAGCAACTCAAGAC CATTTTGTTAGTGTGGATTCGTTGATGTGACCTCTGGGAGTCAGGCTCCTCAGTAGGCTCTCTACCTGGT CAAGCAATTTAGCACACACCCCTTCTTTAGGGACCTCCCAGTAGATCTCCATCTTTGGGAGGAAGGAGAT GACTTCTTGCAGCTCATTCAACAGGAAAAGGCTTCTGAGTGTGTAAATATACACTGTCACTGTTTTTATT TGGAGCAGATTTCATCACAGCTATTTTCTAAATGGCTAAACAAGCATTTCCCTGGGTATGTTCCATGGAA

TATTCATTTTGTAGGAAGCTCAATGATAAAAGAGTCGCATTGTCAATGAGACCAGGAAACTGTATATTTA ATTCCATTTTGAAGAGTCTCAATTTACTTAAGGATAGAAAAGGCACAGTTAAGAACCCTGATTAACTTCT TTAACCCACTTTTCCTAAATGTATCTGCCAGTTTTTGTGTACATAAACCTCTCAACCCTTTGAAGATCGT GCGTTCCAGGGAATACACTCTGGGAAATTCTGGGCAAGACTGTTGCAGGTTTGCAGCAGATACTAGGCAG AGCTGATACTAAAAAGAGGGAGCTGGCTTCCTCATGGCCACTCACCAGTCCTGGGGAAGTCGGGGAGGGA GAGAGGAAGCCTGAGCCTGGAATCCCAGCTCTGCCACTTACAAGCTTCAAATCACTTCACCTCCCTCACC TCGGTTTTCTTTATCTCTTACGTGGAGAATATAAAACCTACATCTTAGTATTCATTTGAGGATTAAAATC AGATGTCAAGCTGTTGAAGATGAGGCCTGGCATGGAGTAGACCCTTCTCCTTGATGTTCCAGGACTGGGG AGATCCAGGGTGATTTTATAAACACATGGGGAGGGTGGGCTGCTGACTAGCTGTGCCAATTCCTAGAAAG GAAGAGTTTCCACTGCTCTGGGATGAGAGATAATCCCAGGAGGATTGAGGATTACCAAAAGGGGGTTACC ACTCCAAAAGGAGAGGGTCCATGCTCGAATGTTTGCTAGTGTGCATGTCAAGGACATCCCTGCAAATCTG AAATCCACATCCTAAATCCTAGCCCCACCACTGACAAAAGATGTGACCTTGGTTAAGTCACTTAACCTCT GCCAGCCTCAGTCTCCTCATCTGTAAAAAGAGGAGTTGGACTACGTATCTCTCTGGAGTACCCCCAACCC TCAGATTAGTGTGGAACCAAATTAAGGTTATTCCAATCAATCCAGGTTGAAGAGTTGGTAATCTAATTAA GGATCAGCTGAATTTTCAAGGTTGAGAGTCAAAAAGAACTGTCAAATAATGTACTTGTGTGTGATGCTTC CAATTTTAATGCTTTTCTGTGTTAGGAGCATATTTTTAAATATTCAAATGTGAATTATTCACATGGCACA TGCACTTATAATGTGTGCATTTAACTAAAGTGAATCATCAGCCTATTATTAATTATCATTTAACCAGCCC TATTCTTTTGGATAAAGGAGAGTGTTAGTAAAAACCAAAGGGTTTGGATCATTTAAGCAAAGCCAGCTCT AAGCAATTTGGCAAGCCCCTCCAGCTGTCCCTCCATATCTTCCAACAGCTATAACCTGTGCATCCTGTGC TCTTTCATTTACATATACATATACATATGCAAATAAACCCATGTTCACTTGGGAGTGTGCACACTCATGT TTGTACACACTGTCCGTGTCCTACCTAATATCCTAGATGTCCATCAAAGTCTTCTTGGGGCAGGTGACAT CCTCACGGGACCTGAGTTTCCTTAGCAGTTAAACACGTGTTCTTTTCCTAGATCTAAGTAAGAGCTCTTC CAGCTCTGACATTTGATGTCTGTGCTCTTTACTTAGCTATTCCTTGGCAGAAGCATTGGAGACATGCTCA TACACATTATCTCACATAATTCTTAAAATCATCCAGGAAGTCTTTTTGCATCCTCAATCCTATTTTAACA GATGATGAAACAGATTTACCTACTTGATGTGTAAACATACAATAATAACTGGCAGTTACAAGCTTAGAAA CCTGATTCTCTATTCTGAATCCAATCAATTCATTCTCCAAGTGACAACCAGGCACTGGCTACTCACTTTT CCCTTCTTAATCAGAGTGGTTTTTGTGGAGTTGGGAATCATCTGGAAGAGCCTCCTAGATCATCTCCCCT CCATAGCCAGGAAAATCTGCTCTAGGCTGAAACATGGGATGTAGGCCACTCATGGGCCAGATACAATTCT AATGGCAATCAAGATATTATAGGTGACTGGAGAGAGTCTGAAGAAGTACATGTCATTAGAGCCAGGGTGC CTGGCACGGGACCAAATGAGCAGTCTAGTTTGAGACAATGGGAAGTTTTCTTAGCAGGAGTCACCCTTCC CCCAACCATTCAGCCATCTGCAGACCAGCCTCCCCGCTCCCCAGATTCCACATGCCTACTCTTTCATGAA TGAAGGCTACAGTGCTATAAGCATAGACTCACACATCTTTAGAGCTACAAAAGACCTCACAAATGATCTA GTCTAGTCTCCTCGCTTTACAACATAGAGAAAAGTGGTATGCTCTAGGACACAGAAGGTATGGTCAAATT TATATTAGGCTTCCTGATTCACTCTTCAGTGTGCTGCTTCCCTGGCCTCTGAAGAGGCGAGGCCCCAGGG CAGGAAAGCACCTCAGTGGCCCACCAGACTTAGCACCAACACATGAGCCTGCCCAGTCCTTTCGCTCTTA TGCATTCCCGTCTCTGGCCTCTCACCCGGTATTTGGGCAGCCTTACCCTTGACCCCTTGTAGCTATATCC TCATTCTTAGATCCTCTGCTTCTTCATTTACTTCTCTGTTGCTGGCCCTGGTTCCCTCTGAACTCTGAGC TCCATGTTAACCCTGTTGCCTACTTTTCCGTTCCTTTTGAAGTAAATTCTGTGTGTCCTACCTCTCCGTT CAACCAGGAACCCCCTAATAGCTTTATGACCTTGGTGTGAAAGTGTCACTTAGTCATGACCCGTCCCTAC AGACAGCAGAATCTGGAGGAATTAGAGCTTAATGTTAGCATTTTACAGGTCACATCCTCTGATCCCAGAG CAATATTAATAATGAAATTATGAAAGCTTTTTTTTAAAAAAAACAATTACAGGCAATTTTAAGAATATTA TTCTAAATTCCCATTGAAATCAAATACGCAATTTTAAAAGATGAGTACAGAGCAAAACTTACTGGATGCA TTACACAAAGTAAAGTTCCATTTCTAAATACTTTCATTTTTTAATCAGAAAGAATAAAAGTATATTATTG AGCACTAAACTTAATAAGTATCCTAAGGAAATCAGGAGAAAAGCAATAAAGAAAAGATAAAATAATAAAT TAGAAAACAGTGGGAATAAAAAAATAAATCGAAGATTTGATCCTTGGAAAATGCCAAAAAAAAATGTGCC AGACATTTGAAGGATATCACATACTCAAATAGGGGAGATTCTGTTAATTCTGTTAATTATTCAACAGAAA GCCTTGGGTAAATTTGGGTAAATATTTAGAAAATATCAAGGCTGTGTCTTACCTCACATCATACATAAAA GTAAGTCCCAAATGTATTGAAGAGTTAAATATTAAAAAGAAAATGTTAAGGGGAAGATGTAGGCAATATT TCCTTGATCTCAAGACAGGAAAGGATATTTAAAGAATGAAATCATGTCTTTTGCAGCAACATGGATGGGT TGGAGGCCATTATCTTGAGTAAAATAACTCAGAAAGTCAAATACCGCATGTTTTCACTTATAAGTGGGGG CTAAATAATATGTACTCATGGACATAAAGTGTGGAATGATAGACATGGGAGACTGAGAAGGGTGGGGGTG TAGGAGGAAGGGAGGGATGATGCAATACTTAACGAGTGCATTGCACACTATTCGGGTGACGGTTACACCA AAAGCCCAGACTTCACCACTACACAATATATCCATGTAACAAAACTACACTTGCACCCCCTAAATTTATA ATTTTTTTAAAAAAGGATTAAAGGCAGGCCAGGCGCGCTGGCTCATGCCTGTAATCCCAGCACTTGGGAG GCCGAGGAGGGTGGATCACGTGAGGTCGGGAGTTCGAGACCAGCCTGACCAACATGGTGAAACCCCGTGT CTACTAAAAATACAAAAATTAGCCGGCACAGTGGCACATGCCTGTAATCCCGGCTACTCAGAAGGCTGAG GCAGGAGAATCTCTTGAACCCGGGAGGCGGAGGTGGTGGTGAGCCGAGATCGTGCCATGGCACTCCAGCC TGGGCAACAAGAGTGAAACTTTGTTTCAAAAAAAAAAAAAAAAGATTAAACGCAAAGAAATAATACATAA TAGGAGTTGATGTATTTTGTTAAATACAAATTTAAACTTCTATATGTTAAAAATCATTAATGAAAAGGCA AATTGGTAATCAGTAGGAAAAAACTTTTAGCATCAAAAGAAATAGGAAAAGATGAAGTAATATCCTTAAT AAATAAAAAGTGCTTAGAAAGCAACAAAAGGGGTCAAAGGACATGATTTCATAGAGAAATAAAAATGATA ATTTTAAATGTTGGTGAGGATATGGTCAGATAGGCATTTCCTTACTCAGAGTTGGACATGTAATTTGGTA CAAGTCAGCAGTAAAGAACTAGAGATTTAAAAACCTTCTATAATCTAATAATGGTACTTTTAGAAATCTA TCCTAAGTCTATCATAAGGAAATAAGTAGAGATAGGAAAAATATTTATATATAAGGATGGTTATAATCAT GTTTATAATAGTAAACATTGGAATCGATAATTTAAAAAGTTACATCATGATATATCCATAATACAAAACT

TACTCATCTATGGAAACTATATTTTTGAAAAATTGATTGTCATGAGAAATTGTTCATGATATAATAAGCA AATGAAAAGAAGGATACACAATGATATAATATTACATCAATTTTATAAATACATATGTGTATATATCATA TATGCATTATGCTTATGCATATCAGTGTCTTTGGGTAGTATGATTATGGTGCACTTCATTTTCTTCTTTA AGCTTTTGTGTTTTCCAGAATTTCCACAATGAGCATATATTATTTATAAACAGAATTCACCAAAGTGTAT TCAAGGCTGGGTACGGTGGCTCATGCCTGTAATCTCAGCACTTTGGGAGGCCGAGGCGGTTGGACCACTT GAGTCCAGGAGTTCAAGACCAGCCTGGGCAGCATGGTGAAACTCCATCTCTATAAAAAAAAAAATACAAA AATTAGCCAGGCTTGGTGTTGCGCTCCCGTGGTCCCAGCTACTTGGGAAGTTGAGGTGGGAGGATTGCTG GAGCCAGGGAGGCGGAGGTTGCAGTAAGCCAAGATCGCGCCACTGCACTCCAGCCTGTGTGACAGAGCGA GACTTTGTCAAAAAAAGAAACAAAAAAAAAAAAACTGGAAAAACCTCACAAAGCGTGTTCAAAATTTTCA AACTTTCTACTTGAAAAACACAGATTGATATTATCTGGATGTGATTTTAAGACTGCTTACTCTAATTAAT TCCTCTAAGAAAGGCATTATTGTTTAATTACATTTCTTAAATCCGTATGAGCTGAGTGGTGTTATTATGG CTGCTCGCCAGAACGTGGTTATTATGCCATTATGTGACCTGGGACCTGTGCTGAGAGGCCACCCAGTGTG TGGAAAAGGGTACTGAACCAGGGGTCTTGGGCCTTGCGACCTCAAGGAAAGTGTTTGGTAGCCTTCTGGG CCTCAGTTTCCTCTTCTGTCAAATAGATCTTACCTGCTTTACTACTTTCACAGAATTGTCTAAAGACCAA ATAAGAAAATCCAGGCAGGAGTCTATGACATGTAGGCTATTATGAAGCCAAAGATCATTATTCAAAGAAT AAGCTAGTACCTGCCTGGAATTCATAGTTCATATAAATGATTTTTCTAAATCTCCCTCTTGGTAAGGAAA CATACTCCCGTGGTTGCAGAAGAAAAAAGCCTCCCCTCTCCTCCCACAGCCCCTTTTCTTACTGAATTAA TAAGTGCACAGCCCTTTGGGGAGGGTGTAGACAGATCAGGCACACCCCTGTGTCCACCTCCACCAAAATT CTAGGGGAAAATGCCTGGCTTCTTAGGAAAATGCTTCCCTAAAAAAAGAGGAGAAACCTCAAGTGTTCCC ATGGAACTGGGGCAGAGGGCAAATTCCGCACAACCTCTGCACAAACCCACGGGCTGGCTGCTTGGCAGGC CTACAGGGTCCAGATCTCTGATCTGTGGCTCAGAGTAACGAGGTCATTTGGATGGTACATTGGTTTCATA AACCCCTCCATGATGCTGCTGCTGTTGTGAGGATAGTTGTCACTTCCAACTCATATTATTGCAGACTCTA GGCTACACGTAGTTATAGGGACAAGAAAGTCCACAGCTCCTCCACTCCCCACACCAACGGCTCCAGGCAG ACTTGCTGCAAGGAGCAGAGACCCCTTGGTCACCACAGGTAAAGGGGACAGTGTTTTGACTGACTTGGCA AGCTTACCATGTGCAACCCTGGCTGCATATGAGTCTCCTAGGGAGCTTTTTAGTAACCCAGGTGACACCC CACGTTTAGAATTTAACCAGAATCTCCAGGAGTGGGAACCAAGTATCAGTATTTTTAAAGCTGTCTGGGT GATTCCAAGAGCAAGCAGGATTCAGAAGCATTGGCTCAGGTTTCAACACAGATCTAGGTATCTCTCCCAC ACCAGTCTTCAAAAAACTTATCTTGGTGGGTTTTCCACCACAATTCTACACTCTCCTAAGCATTCTTTGC ACGTAATTTTACCTACCATGGATCCAATTCAACCCTAAGGCATCCTTTATCCTTTAGCTTCTCTACCGGC TGGCTCCATTTCTCAGTCTGCCAATTCCAGATCCCCAAGAAGGGTGTCGTCTTAGCCCAACTCACCTTTT TAAACCAGGCTACTTGGGCCATGGGATGCTGGTTGATTTAGAGACTGGCTACTCTGTGGAAAGGGACAGA GGTTGTGCAGTGTAGAACACGGCCACCCTGGTCTGCCCCAGTTCTGTTTTTTGTGTAGGTGGGGTGGGTT GGGCACGGTTTCCCTTAACAGAGACTGTGGAATACTTTAACATATCTGGTTCAATAACAAATTATAATAA TGAAATCAAACATATTTAGAGGAATCCATACAAATTAAGGGTCAGAGTTTCATCCTCTGTATAGAATATG ATGTAGGAGTCTTTTTAAGTATACGTAGGATAATAAAATGGATAGCCCACTCAACTATCTTTAGAATGTA TGTATAATTTTATACATCTTTTTCCATAGAAACTACAATTGAATTTAGCATTGAGAGATTATGGTATTCA TTATTATCTTAGCCAGAGGGTGTTTTTTTAGCTTTTATAAGAAACTTTTAAATTGAAGTATAATATACAT AAAGAAAAAGTGCACGTTATAAATGTACATCTCAATGACTTTTTACAAAGTGAATACCCTTGTAACCAGC ACCCAGGTCAAGAAACTCAATATTATCATTACCCCAAAAGATGCCCCTTGTGTTCTCTTCCAATTACTCC TTGCCTCCAAGGGTAACCACTCTCCTGATTCTAGCAGCATAGATTAGTTTTTGTACTTTATATAAGTACA GTCATATAGGATGTACTTTATTGTGTTTTGGCTCCTTTCCTTGAATATTCATCCTTAGTTTTGCATATGC ATTTGTAGTTCTCACCAGAGGTTTTAGTCCTTGAAATGTAGTTCAACAAACATTATTCAGCCAGGAACTG TGCCCGGGGCTGGGGACACCAAAACAAATAAGGGCAAATTCCTGCTGTAAGAAATATTTAATCCAGTAGT CTCGTAATGATCAGAATTTAGTGTAAGGTGAATTCCTCTTTGTCCAACAAATATTTATTCAGTGTCCACT AGAGGACAGGCGCTGTATCACAGGTGCTAGGGATAAGAAATGAACAAGCAGACAAAAAATCTAGCAGAAA TCCTCTCTTTTCTTCCCGTGACGTGCATGGGCAAGTTCTGCCCACTAATATTCATTTCCCACTATGTCCT GCATCTGATTTAACATAAAGGGAAGTAGCAATGGAAACTATTTCTTCTGGCAACTTGGAATATGAGAAGT CCCTGGTCCTTGGGCCATTTTTCAAATCAGTGAGTGTGGGAGGTATAATATTTGTGTGTACCAATTTCTT CCTTATTCCATTCTCTCCACTGCTGATGCTGCCTATACTGAATGCAATGAAAGTCTGGAACCTAAGGAAG CCTTTGGGTGTTAGCTTACCTTGGAGTCAAGCTGAGCAGTCTTCTTGTTTTCCAGAGCCATGAAGTGAGA AGAGACACAATTGCCAAAGCTGTGATGGAAATCATACAGCTTTGAATTTGTTTTAGTAACATGGAATTCT AACATGTGTCACTGTTCATTAGTACTGGCTTTTCAGAGCCTCGTGAATTACTTGGGTACAAATCCATGAC ATTAGAGGCATGAGGGATTAGAAGACAATCCAACACCACTCTAGGATTGATAGTTTAGGAAGAAATGCAA GAAATCATCTCATTTTGTTTGGTTGGTTCATTCCTACCCCCCATTCTCCACATTTTTATGAAAAATTTTA AGTATATATAAATGTAGACAAAATAGTAAAATTCACTCTTATGTATCCATCTTTCAGCTTCAAAAATCAT CAACTCATGCCAATTCTTCTCTAACCCCTTGTTGTCCCCCTCCTCTGATTATTTGAAGCAATCCCAGATG ACAAATAAATTACTCATAAATGTTCACAAAGTGAGATCTCCTAGTTTTAATTCCTGTAGTTCACAGGTTC AGCGGTGAATTCACTAAGCTAGTTAGTGGTACTTATATTTAATCCCTGGCTTCAGAAGTAAAAGTGATAA GCTTTACTAACAGAATTACTATTATTATTATTACATTATGACATAATATAATATATTCTTGTACTGTTTG GCCTAAAACTACCTCTTTTAAGCCTCTCATTCAGTTCTCCAAGGAGTTCAACAGGGTGTGGGATGGCTGA TTGATGTTCTTTTTCATCAGTAATTTCTAAACCAAATCCTTGTGGATTTTGAAATTGCAAATATATTTGT CCAAGTAAAAGCTTCAACTATTTGGTAATAAAATGGGTCTAAGACTCTTCCCAAACCTCATGTTTCCCAT TTTCAGTCTGTGTGCAAAGGAGGAGGTTGGCTGTTGTTCATGGAGCCAGTGTCAACTGCCTGTCTGTTAC GAGATGTCACAGCACAACCAGGCATGCAGCTTACTTTTTTTTAGGTTTCCCAGGCTCAAAACCGCAAAGT ACCATTGTTTTTGCCAGTTAGAACACTCCAGAGGTTTCTAGAACAAGCAGAAAAGATTGTTAGTGCCTTT

GTCATGAAAATATGTAAAATGTTGCTTTGGCTGCTTTGGCTATCATAGCTGCTCTGATTGTATCTCAAAA TGTCACCCAGGGTATGTGGCAGTCATTGTGAGGAATCGGGCACCAGGCACCCCTTGATGAGCCTTTGATG AGGAGATGATTAGGAGTAAAAAAATGCTGCATAGATTATCTGCATAACTAACACTGTAGCAGACAGCCAT GCAGACATTTTTCTGTATACAACTCCCACTCATTTTTCTGAGAGATAAAAGCAGCCAAGATTGACAATCT GTCAGTTTCACCATTTTCATTGTGTACTTTTTCTGGGCAGGTTATTCACATCGAACTCCTTTGATGTGAT CAGTGATGATGCTTTTATTGGTCTTCCACATCTAGAGTATTTGTAAGTAAAAAAGCTTTTTTAAAACATG ATGATTCAGGACAAGTACTCTCAAGTTCCTGTAACTGTTTGACAAAAAATATTATAACTTATTGCAGATT CATAGAAAACAACAACATCAAGTCAATTTCAAGACATACTTTCCGGGGACTAAAGTCATTAATTCACTTG TAAGTATGAATGTTGCTATTACTTTTTAAGCTTGCTAATGGACAATGCAGTTTGATCACCTGTGGTTAAT GCACCTACTTTATGAGTGTCCATAAAAATTTAAGAAACCCAAATGACTTCTCTCCATGCACACTTGCCAT CTTTCCCTGTCATCTGTCAGAGGGTGTTCAATAAATTGATTACTATTAATATTTTTATCCAAAACCAACC ACAATTAGAAAAGCAATCTAATGCAGGGAAATTGTGTGCCTGAAATAACTTCTTTTATTATCTACATCAT TACTTGCTTCATGGATCATTATGAGTTGACTGTATAATTTGATATCAGCTTTTTTTCCCTCTATAAGCCC TCACTTTATATACATTTAAACATAATATAGCCACAGGCGATGTCTTCATAGTACCTGATCAACTTGTTTA TCAGTGAAATATTTAATTACTCAATGGATGATTACAGCTAACAGCAGTTGTTCTTAACAGGGAACAATTT TGTCTCCCAGGGAACATTTGGCAATATCTAGAGACATGTTTGGTTGTCACAACTGGGGATGTAAATGGCA TCTAGTGGGTGAAAGGCAGGGATTCTTCTACTGCTTAGGACAGCCCTTCACGACCAAGAATTATCCAGCC TAAAGTGGCATCGTGGGCAGGTTGAGAAACCCTGAACAACAGGACCCAAACAATTTGTTTCTTCTAGCCT CCCTCACCCTCTACAGACAAACACATTCACACACACACACACACACACACACACACACACACACAGGCAC ACATGTGCATACACACACACACTCACACACACACTCTCACATACACACAGATTATTTTAAGGAAACTAGA AGAGCATGGAAAGAAAAGGCACTAACATTTACATCACTTTTCATCTAATCCTCTCAATAATCCTGCCAAA CAGGCACAATAATCCCTGTTTCAGAAATGAGGAAACAGTCTTGGAGGGGATATATGCCTCGTGAGCTGAT TAGTTACGGAATTATGAGAACATCTACTTAATTGTTGAATTGCAGTGACTCTTCATCCTTTCAACAGCTA TTTCTCAATCTACCTAATATGTATCTTATCAGTTCTAGATACAGATGCAAGAGATCCAAGTGTCATAGTC ATTCTGTCAGCTTTAACCTGTTCTCTCATCATACTTCATTCCCAGCTCTGCTCTAGGCCCTAGGATGCAG CAGGGAACAAAGGAGACAAAAATGCCTGTCCTTGGACTGAGCCTGATTTCCAGTGTGTCCACGCGGTTGT CACTCTGCCTCCACCAGGCCCCTTTCTCTCCTGTTGCTTGTTTTGTAGCAAAGAGAAGCTAATAGCCAGA TATATTTTACAAAATCATGCCTTTCAAGGCTTCTGTATAGAGTTATTGTAATAAAAAAACAGTGTTTGCA ATACTACTCTTCATGTCAATGACATGATATGATGCAAAGATGGCTGTTTACAGAATGAATGCTATGCAAT TTTATCACAGAGCTCAACACTGCCAAACCTCATTTTTCAGACAACAAATAAACAAAAGAAACAAATATAT GGAAGGAGAAATGGAAATTCATGACCTCAACTTGAAATCCACTAGATTCACATACATCCCAATCCACATG GCCTCTGTCCAGTGCTTAGCATCATGAGAGCTGTAGGGAGGATGCACAGCCCAATTCATCCAGCTGTGTC TCCTCTCCAGCCTAAAAGCCTGTGGTTTCTCTCAGGCTCCATGCACAGAACAGGAAAAAATGTTTCCCTG TCTTGGAAGGAAGGAGGGAAGGAAAGAAAGAAGGAAGGAAGGAAGGAGGGAGGGAGCGAGGGAGGGAGGG AGGGGAGGGGAGGGGAAAGGAGGGAAGGAAGGGAGGAAAGAGGAAAGGAGGGAGGGAGAAAGAAAAAGAG GAAGGAAGATGAGAGAGGAAGGGAAGGGGGAGAAAGGAGGGAAGGGGAAGAGGAAAAAGGGAAAGAAAAA GTCTGTTTGACATCCTAGTGTAGGACACCCATTTTGTTTACTGGAGCCTATGATGTGTATCAAATGAAAT GGGAATTTAAATATGGGTCTTAATGATGCCCTTTCTTTAGCAGTATGGAGCCTGTAAGAGAGAGAGACTG ATACAGAACACGCTTAACATTCAGAAAATTTCAGGTTCGAGGAACACCAAGGGCAGCCTTCAAAGAGAGG CATGACTCTTAACAGGCAGTGAAAGAGAAGTCCCACCTCTCTGAGCTGGTGCTGGAGGTAGGAAGGTGTT ATGAGGGAACAGGGGTTAAGGCTGTGGCTAAATGAATATGACCCTTAGGTCTCTTGTATCTTTAGCTAAC ACTTATTAGGTTGACTGATACGGCATTGCCAATGTTAGGCTGTTTTTTATCTACAAAAATGGCAATGCCA TATGGTTCATCTTAAATAGTTTCTAAGATGTGACACATGCTTTACAAGGAGTAGGGCCTACAAAGATGAG TAAGATCTAGTCCCACCTTAGAGAAGCTCAGGCTAGCAAGAGAAATTGGACAAATATTTTCTCACTGAGT GCCTTTGTCCCCAAGTTATGTTCTCCTTTCAAATTATGCTATTCTTCATGGTACAATTCCTTCCTTCCAT CCTTCATTCTTTCCTTCCTTTCTTCCTTCCTTCCTTCTTTTATTCCTTCCTTCTTTCCTTTCTGTCTTTC AAGGAGAGATGGAGTAGGGAGGATCTGAAACCCAAATTTCCCTCTGCCGGCTCAGTAAGTCAGGTTCATT TTCCATTATTTATGGCGTATTAATAAATCTGGCATTTGTCTGAGCCATTAGTTTCCTTTTTAAGACATTA GGCTTCAAAGTGCACTTTTCTCCACCTCAGATCAATCCTTCCCAATCAAGGCAAACATTCAAGCTGTGAG GAATTAGTACCTCACCCTCTAACCTGCATGCATTGCAGGAAGAGGTTACAAAGCTTTCTCATATTCTTGA CTTCATCATAGTCTCCTGACGACCCCGTGAAAGAGACAGGGCAGTAGCACTAGGTTCATAGATGAACGAG CAAGTGGAGGTATACCATGAGTTGAAGCTGCTCCCCAGTTACTAAGTGGTAAAGCCAGACAGCTCCCACC GTCTTTGATGAGACGTCCAGTGCTACGTCCACTGTGCCAGTTACCGCAGGACCAGATAGCTTTTATTGAG CTGGCTGGATCCTTCCATGGGACATGATTTGAGGAGAAAATATGACTAGAGAAGAAAGAAAAGATCCTGA AAACAGGAAACCTACCCTCATATTTCATCTCAAAATGCAGATAGTTGGGATTAGACGTATACTTTCAGGT CCAGCTGGTTCCCACTCTGGCCTAACATTAAAGCCACTCCCTTACCTGCGCCTTTATATAAATTTAGTTC ACGCCTAAGAAACAATGCACTTTCTCACCTTCTTATACAGAGATGTCAGTAGATTTCTCTTCTAAGAGAC ATTTACAGATAACTTAAAAGTTATTATAGGCCGGGCGTGGTGGCTCACACCTGTAATCCCAGCACTTTGG GAAGCCAAGGAGGGCAGATCACTAGGTCAGGAGATCGAGACCATCCTGGCTAACATGGTGAAACCCCATC TCTACTAAAAAATACAAAAAATTAGCCGGGCGTGGTGGTGGGCGCCTGTAGTCCCAGCTACTCTGGAGGC TGAGGCAGGAGAATGGCATGAACCCGGGAGGCGGAGCTTGCAGTGAGCCGAGATCACGCCATTGCACTCC AGCCTAGGTGAAAGAGCAAGACTCCGTCTCTAAGAAGTAAATAAATAAATAAATAAATAAATATAAAAAA GTTATTATATATACCTGGCTATCAGGCGAGATCCTCCCCTCCCCTGCAAATTAACCCTTAAAACCCTGCA AATTAACCCTTAAAAAGGGAGAGAGACTTCTTAAAGTTAAGTCCTTTCAAAGCCTGTCAAATGTTGGGAT TCTCAATTACTTTGTTTTGAATCATTGTTCTAATGTTATGCATGTTTCACTTTAATGTTTGAATGGTTTG

TATATTTTAGTAATCTTTTGTATATTTGAATTTTTCTGATGAATATATGTTATTTTGGTAATCAGGAAAA CATGAATAAAAAATTAGTTTATGAATAAAAAATTAATTTATGGCCAATGATGACAAAATCTTCAAACATT ATGTATTGGCTTCTAGTAATACCTATGAGTCTTTAGTAATGATTGCTTTGTGTGCCATTAAGTGTCACTG TGGCTAAGGCCCAGTGGAGCCAGCATTGTTTGCATGCATATATATAAATATTTGGAGAAAATTATCATCA ATCTATATGTATTGTAGAGTATCCAATGGTTGCTCAGCACTGAGGGGGTACAAAGGACTATATAACAGTC CCCAAACTCAGGATGCTTGTGTGTTCCCAATCAGTTGCACAGGATGTCTTTGTTTTAGCCTTCACATTGC CTTATATGTAAAGAGGTAGTGCAATGTAGCAGCTAAGAGGGTGATCTCTGCCACCAGACTGCCTGGGTTC AACTGTCTCTCTGCCAGCTCAATAGGACAAGTTTATTTTCCATTATTTACTACTTATTAATAAATCTGGC ATCTATCTGAGCCATTAGTTTCCTTTTTAAGACTTTAGGCTTCAAAGTGCACTTTTCTCCACCTCAGATC AGTTCCTCGGCTTTGCACCTTAACAGCTGTGGAATTTGGGGCAGGCCATTTAGCTTCACTATGCTACAGA CTGGGCATGTAAAATGGGGTTATAATAATAATCTCTTAAAATGGGATGGGGAATAACAAGGCATTTAAGA GTATTAGTGCCCAAGGTTGTCTTGAGATTAAGTCAGTTAATAAATGCAAAGTGCTTAGAACAGGGCCTGG TATGTAGTAAGCACCACTGTATAACTAAAACTCACTGCCCAACCTCCATTAGTCCTGGATCAGAATTCTA AATGCTATAGTGTATATACCATTTAAACATTTTCTGTGAAATTTAAGTAACTACAAAGTGCTCAGAACAG GGCCTGGTATGTAGTAAGCACCACTGTGTAGGAAAAAACTAAAACTCACTGCATAACCTCCATTAGTCCT GAATCAGAATGCTAAATGCTACAGTGTATATCCCCTTTAAACACTTTCTATGAAATTAAAGTAACTGCAC TTGCACCGCCGCAGGTTCAGAGAGACACAAACCCAAGCTGAGAATCTGCATTGCAGGGGGGCTGGTGCAC CCCAGATGGCTGTGGGCGATGCTTTCTTAGCAGCAGAGCCCGGTGGCACTGAAACAAACGCGGTGTTTGA GCCAGTCTGGTCTGCCTTAGAGAGAAGACTTTCCAGTGGGTTCCACAGTCACAATCGGAGGTTTTCTTGG GTAAATGATGAAAACACCAAAGGGCAGGAAATGGTCATCATTTCTGTGAAGAAGGAATGGCCGTCTGCCT TTGAGAACACTTGACCTTACTCAGCCAGTTTCAAGGTAACTAGACATTCAGCTGCGCAATATCTCAGTTG TTTTAAAGAATGTTTCTGGCCAGGTGCGGTGGCTCACGCCTGTAATCCCAGCACTTTGGGAGGCCGAGGT GGGCGGATCACAAGGTCAGAGATCGAGACCATCCTGGCTAACACGGTGAAACCCCATCTCTACTAAAAAT ACAAAAAATTAGCTGGGCGTGGTGGCGGGCGCCTGTAGTCCCAGCTACTCGGGAGGCTGAGGCGGGAGAA TGGCGTGAACCCGGGAGGTGGAGCTTGCAGTGAGCCGAGATCGCGCCACTGCACTCCAGCTTGGGTGACA GAGCGAGACTCCGTCTCCAATAAATAAATAAGTAAATAAATAAATAAAAGAATGTTTCTTAGGTGAACTT TGAACCAGGACCTAGAGAATGGTGCTTAGTTGAAGGCAACTTTAAAGAGCCTCCACGTTGCCTTGGGATT TCTACCACAGTGGGGCTGTGGGGAGGGAGGGTCCTTCCTGAGGTGGGGGTCCACACCAGCTGGTGGGGAC TGCACTAGTCACTCACAGGGCTTTGAGCTTTCTCCCCCAGTGCCTTGGCACAGGGGGCCTCAAAGGAGCA TCCTCTGCTCTTTTCTTTGCCTAGATATGTACGCGTCCTCTCGTCTACTCCAGCCTGCCTCATTCATTCA TTCATTTAAAAGTATCTATTAGGCTGTGTGCGGTGGCTCATGCTTGTAATCCCAGCACTTTGGGAGGCCA AGGCGGGCGGATCACTTGAGGTCAGGAGTTCAAGACCAGCCTGGCCAACATGGTGAAACCCCATCTCTAC TAAAAACACAAAAATTAGCCGGGCATGGTGGCGTATGCCCGTAATCCCAGCTACTTAAGAGGCTGAGGCA GGAGAATTGCTCGAACCAGGGAGGCGGAAGTTGCAGTGAGCCGAGATAGCGCCACTGCATTCCAGCCTGG GTGACAGAGTGAGACTCCATCTCAAAATAAATAAATACATAAATAAATAAAAGTATCTATTGAGGGCTTA CTTTGTGCCAGGCACTATGCTAGCACTACAGATAATGGAAAGAGAACCATCCTTCTAGTACTCATTGCAG AATTATCCTAGAAATAAAAGAACATTGATTAAATGTTTCCTCAAGTCTCTCATTGACTGTGTCTTCACCC CTCTCCCAATCGGCTCCACCCCACCCCAGTCAGATATTCTAGTGCCACCTAGTGTGACCTCCATGCCCTG GAAAAGCCTGTTGTCAAACTCACTGACCCCTAAGTACGTGGCCTAAAGTATCACCTATCCTAGTGTCATC TATTAATTTTAAAGATGAAGCCATAGCCTCTCAGTGAAAATGACAAAGCTGTGATTTAAGAGCAAGAATA GGACTTGGAGATCATTCTGACCCAGTGATCATTCACTTTATAGAAAAGGGAATAGCCTGAGCTCAAACTC AGATATCCACTTCCCTGTCCACGTCACCCCTGTACTCTGTGGCTGAGTTCTGGGCCCTAAAGTGAGTCTG ACTGGCAAGGGGTCCACTTTGGCCAACACAGGAGCCTGGTACTCCTTAAACCAAAACATGTAGAAAGTCT TTTTCCTGACAACTCTGGTGGCTACCTGGGACTCCTTGCCTCTCTGTCCTTCATAATTTTTCTTCCTCCT ATTCCTAAGAATTAAGAATTCTTATATCTAGTATTAAAATTCAGCTCTGCTTAACAGCTCTTGGCTAACA GATCTTTGCAGAGGAAGGAGGGTCCCCTACTGCAATTCAGGGTGACACTCTCTCAAGTCGTGACACTCTC TCTAGTCATGACACTCTAGAGGAAAGAGAAGGACTAGACACTCTGTGGGCCAGGATCCATAACATGATGT TCCAGAATCACTTGCTTTTGATGGGCTCTGTATTAATGGCTGGAAGAAAACATGACTAAATCTTAACAGT GGTTACTTCTGCTATGATAATTATTATTTCTCTCCTTTTTTAAAAAAATAATTTTTGCACTTTCCAACTT TTCTTGAAGTGAAGAAATAATTATTATTCCATATATATGTGCATATGTAGTGTGTGTATATGTGTGTGTG TGTATCAATAAAAGTAAAAGATGGGACTTTTAAGAATACCAATGCATAGGATTATCCCTACTAAGAGAGG CAGAAAGAATAGATGATCTCAGCTTCCTGAAGCAAATATTTTAAAGTGTTTAGATATTTAATATCACTGT TTAATGACAATAACAACAAAGAAAAAGATTAGTTTTGTGGTCTTAACACATAAGGTTTAAAAATTCACCT GCAGATACAGTTTTTCCTGGAATCCATTACAAAGATATGTATATAAAAATATTAGTAGCTACATGTGTTA ATTCTCACAATAAGTGAAATTCATATCCTTTACTCAGTTTTGCAGATGTCAAAACAGAAACCCAGAGAGG TGAAATTAACTTGGCTAAGGACACATTCCTAAGAGTGATGACGTTCAAATTTATTCCCTGGTGGATTTGG ACGAAGTTGGCACTTAAACCACTATACTCCCCTAAGTGATGCAGGAAAGAATAGCTACAACCAGTTTTAA AAATACTTAGCACGTATAGCAACTAGAGCTTCTGTTGAGGAAAAGTTAATAATAATATTATTATATCATT ACTCTGTGAAGCTGGGAGTGGGATGAAGACAGAACTGGGAAAATTTTATCCAGGTATTTTGCTTTCTGGT AGCAAAACTCGGTGCAGAGGTGGAATCTGAAGAATCAAATAATCCTAATATCAACCCTAAAATAACCCTA ACCCTAACCCACAAAGTGCTATCTCAGTCTCTCTAGCTGTTAAAATTCCCTGATAGACACAGGATCTCAT TCTACTTACCCCTTTTTTAATGACTAAATGCTGAGGAGGCTTCAGGGAACGTCGCGTGACTTGCCCTTTT TTTTTCCTGTTGCCCCTGAGCCCGGTGCCCTCAGCAGCCCTCCGCTGCAGCAAATCTCCCTGAAGCCCTC AGATGCCCATGATGATTTGTGGAGGCAACAGGCCCTCATCACAGCTGGCTTTAAGCTGGCGCAAAGAACC ACAGCGGTGGCTTCAAGCCTCGGCTGCCCTGTAGCCCCAAACCTCCCTAAAGGGTGAGGGCCTGAAACCG

CATGAGAGTTTTCTAGGAAACAGGCAGCTCCCCGCTGCAGGAGGTCTTGTGAAAGACATTTGCATGACCT GACCACTCAGGCCCCCGAGTTCGGTAATTCCTAATCTAACCCTGTAGGTAGCTTTGGAACTTGACAAATG CAGCCAAACGATTGATTTCGAATGACTGAACCCTCATTGGACAGAACCATAGATATATACAAATACTAAG CAACAGGATCCTTTCAAAAAGTCAACTTTATTCTTTGTTTCTTCTGATTATAAAATAATAATACATGCTT GATGATAAAAACTCACAACATAAATATGTATTTACACAGAAAAGTGAAAGATTCCAGTAACCCTACACCT ACCTCATTTTTAAAATCTCCTACAAAATATTCCGTGTTACATATGGACCATAGTTTATTTACATGATCCC TTATATCTCTTTGTTGTTGTTGTTGTTGTTGTTTTTGAAACAAGGTCTCACTCTGTCACCCAGGCTGGAG TGCAGTGGTATGATCATGGCTCACTGCAGCCTCAAGTTCTAGGCTCAAGCCATCCTCCAACCTCAGCCTC CCAGAGTAACTAGGACCACAGGCACACACCACACCCAACTAATTTTTGTATTTTTCGCAGACAGAGGATT TTGCCTTGTTGTCCAGGCTGGTCTCAAACTCCTGACCTCAGGTGATCCGCCCACCTCTGCCTCCCAAAGT GCTGGGATTACGGGCGTGAGCCACCATGCCCTGGCAACCTGTTACATTTTTTGCAGAGTAATTCTCAGGC TCTGTCAGCATTGCTTTCCTAAGTCGTTCTCTGCCCAAATCCCTCTGCCTACTTTCTTTTTATCTAAATG TAACCTGTCACTGGAGCCTCCCAGAGACTTAGGGAAGCTGTCTGAACTCCAGCTCCCTAGCCTTGGATGT CTGGAAGCCTCACTCTCATTTCTGAATCTTGCTTCCTCTTGAGTCTGCTTCTCTTAGTGAATCCAGGCAG CCTTCTCATGCCCATCACTCACCAAACCTAGACATGAAGCTTCATTCTCTCTCCTCTAAGAGCAGTTCAT TGTGGGCAAGGATTTCATTGAATTTTTGGTAGCGGAGCATCAGAGGGTTGACACTGGGTTCAGCGAATCC ACCCACCATTTCTCCCTCTGGCTCCCCTGCCTCCCCCACCCTCTGATTCCCAAGGCAAAAGCCTGTCACA AGAGGTTCTCAGGAAATTTAATGCCTCATTCCACAGCCCATCGGGCCAGCCTGTCCTTCTCAGCCCCATC TCACAATTCTGGTGAATTAGAAGCCCTCCATCTTGTCATGCAAAAATACTGCCTCTCCATTCTGGAAGCC CCATAGCTCCAAGTCTGCAGCTCTGTTTTGCCATCTGTTCTGTGGTTAGCTTGCACGTGGATCATGTGAG GCCTAAAGCTCTACTCCTTTGGGCAGGAAGTAGAACTTCCAGATTCACTCATTATTTAAAAGAATCCCAC AGGGTACAGTGGCTCACACCTGTAATCCCAGCACATTGGGAGGAAGAGGCATGAGGATAGCTTGAGCCCA AGAGTTCAAGACCAGCCTGAGCAACATAGCAAGACCCTGTCTCTACAAATAAAAAAAACTTAGTTGGGCA TGGTGGGACACACCTGTGGTCCCAGCTACTGGAGAGGCTGAGCCCAGAGGATCACTTGAGCCAGGGACGT TGAGGCTGCATTAAGCTGTAATTGCACCACTATATTTCAGCCTGGACAACAAAGTGAGACCCTGTCTCAA AAAAAAAAAAAAAAAAAGAATTCCCCTTGGACAGTAAGACAAGCATGGTGCCTTCTCCTGTATAGCCCCT ATTGGCAGTAGTAATTCAGGATTTGACTGCTTAATCATTGCATGCCCACTGTCCTGTGACTCAGCCTCCT CTAAATTTATGCTCTGCAGCCAGTTGCAGTTTTTACCAGCTTCAGAGGAAGGGACATCCACTTAGAAGTT AGTATGATAACTTTAAGCCACATTGTGCCAGAAAATAAACTGAGACACATAACATGGGAGTGACTCTGGA CTGGTCCAAGACATCAGAAAATCTCTTCATTCAAACACCAGTCAGTAAGATGAATTATGGAGACATTCTC TTGAATAGTCTTCCTTAGCAGCATTTTATTAATTTTGCTGTATGTGGACAGCCATAGCAGTTCATTCATG TGGTCTGTTTCTAAACGTTGTTTCTTGGACAACATACGTACAGAAGCTGTTCTCTTCTGACACCAGCCAC TCTACTGAGAGTAGTTCAGACACCTGCACCATCCTTGATTATCTCATTTCCTTTTTTCAATAATCTTATG AGATAAGGTATTATTATCTTCACTTAAAAATGAAACAACTGAATTTCCAAGAAGATACATTACCAAAGTC ACACAGCCAGTAAATGCTGGAGTCAGGATTCAAACATAGCTCAGAATCACTCCAAAAGCCATGCTCTTTG TCACAATTGGACCGTCATTGACAATGGAACATTCTGACTCAGTTCCTATTCTTGCATTCCAAGGTTCATT GAGAACCATTTTCCAAACTAGCAGTAATTTCTAAAACCTAGAAAAACAACATAACTAACTTAAAAATAAG TTCTAAAGTTCATGAATGTTTGCACTTGAGAAACCTTATATTTAATTAGTGGAGGTAGAAGACATAGACT TTTGAGAATGTTTTAAAGGGAAGTTGAGAGAATTTTCAAGGGTTTAGAGGCTAATGGTATGGGCTCTTTA GTTTTAGTTTAGTTTTTTATTTTTATTGTGGAAAAAAAAAACACCTCTAAGAAAAGTTTATAACGTGTAT ATATATGGTGCTGGTTAAAAATAGAAAATTACACTACTTGGTGACCTTCCCCAGTCATACAGCATTAGCA CTGTGCTATGTCTATATTTGTGTACTAGGGACACATTTCCCTTTCCTTGACATTATCAGGACAATTTTTT ACTCATTAACCTTTATTAGAATCATTCTAGTGATTTACGGAAGAACAAATAGAAAGGGAAAGGGAAAAAC ATTCCTGTGATCACAGGAAAAGAACCAGAAGAAGGAGAAAGAGGGGGATGATTAAAGAGAATAGATGGAT AGGTTTTTATAAGAAAACTTTTTTCTGAGGTGGAATGAATGGTGCTAGAAAGGAAAAGGAGGCGTGGGCT TTAGAAAGGAGAAAGGATGTACTGGGGAAGAGATAGAGCTACTGGCACTGTGGGTTGGAGGTTTGGTTGC TCTCCGGAGCAAGCGAGGAGGAAGCAGGCAGAAGGGGCAGCCCAGGCAAGAGGAGGGGGCGCATTTTGGA GATGGACACTCACCAGTTTGGTTAGTCATCATTTGGGAGATATCCCCTAGATTGAGCGTCTGAGAGTGTG AATCTCTCGTGTTCTGTTCTAAATGGAAAGATCAATGCCCAGGAATCATTGTTGTTTTTAACCAACATAA AGTGTTGTTTAAATATTGAGTGCACCTGTAATCCCAGCACTTTGGGAGGCCGAGGCGGGTGGATCACTGA AGCTCAGGAGTTCAAGACCAGCCTGGCCAACATGGTGAAACCCTGTCTCTACTAAAAATACAACAATTAG CCGGGTATGGTGGCGGGCACATGTAATCCCAGCTACGGCAGGCTAAGGCAGGAAAATCACTTGAACCCGG GAGGCAGAGGTTTCAGTGAGCTGAGGTCACACCACTTCACTCCAGCCTGGGTGACAGAGCGAGACTTTTG TCTCAAGAAATAAAAGTAAATAAATATGGAGTGGGTGTTGAAGTGAACAGGTTAGGAACTGAAAGGACAA TAGCAGGAATCTGAGTAGTAGTTCAGAAGAAAAAAAAAGGACCACAAGTCTTTTAGTAGGCTGGAAATGA CAAAAGAGACTCATCACTAAGCTTTATCATTAAATGCCTTTGTTTAATTTATCACTACAGTTTACATCAC CTTTTTTTTTTTTTTTCCAGGAGCCTTGCAAACAACAATCTCCAGACACTCCCAAAAGATATTTTCAAAG GCCTGGATTCTTTAACAAATGTGTAAGAGGACCTAAGAAATCACTGAACAATTAATTAATTTGCAGTCAT TAACAAGGAAGCCTGGTATTGATTATTATAATTTTTAATTGGCTTTTAAATTAAAATTAGGATTTTTAAA TCTAAATTTTAACTGGCCAAAAATTTAAATTTACATGGACTAACATTTGAGTCTTCTATGGTATTTATCA TTGGGAATTCTGAATACATGGCACCCAGGCATTCCTTTGACAAAGTGTGTGTCAATACGGCAGGGAGCGA TGATAAAGGACTTTGACGGGGTGGGAAGAGACCCGGCTAGAAATTTCAAAATCCCTTTTTTCTCATGAGG TCCAGGAAATAGGGCTACAAAGGAAAAACATAAAGGAGTCTGGGAAACTGACAGAATAAAGATGAAAGGT TACATCCTTGGGTTGGTCAATCAACAGAGAAATGGAACAGGAAGTGGGGGAAGGGAGAAAAGTTTGCCTT TTTTCTCATTAGACTGAACAACAACTCTAAGCTTAATCCCCAGGAATTTGGCTTAAGCCCCAGGAAGGGA

ATGAACTTAATTATTTTTGACCCTTTTTCAAACAAAAGAGCTTTCAAACTTTGACTTTTAAGCCTCCGAG TGAAAGATCTTTGCAAGGAATTTGTATATTTTGCTGTAAGCAAAGCAAGTAATATATGCCTAAACTTCTC AAAAGCAATTCAGCTTCATTATTTGCCTTCCAGAACATTAGAAAGGTATTTAAATGTCATAAGGTGGAAG ACAGGAACAAATGAAAGGATATATGAAAACAAAAGCTTTTTTAAAAAATTATTCAGGTACTTGTCTGTAG AATATATATTCAAAACAACTTTTCTCCAATAACAAACAAAAAGAAAATCTGAGTCAAGTACTTTCTTCCA AGCCCAGTATTTAACTTCAAATCAGTAGCCTAATGAAAAACTTTCTTTGTTCCTTGCAGTATTTCCCCAG CCCCTCCTCCATGTCTATTACCTCAATTCCTGAATGATATGGTTTAATTCCAGGAGAATGCCCAAATCAG GATTCTTCCAAGAGAGTCCTGGACAGACCATAGTTATCCTCTCTGGTTTACAGTAGCCTGAAAGGTTGCC ACTCATTAGTGTGGGTCAGAGTTCTCTTGGGAGACTAGGTCAGTTCTGGGAGTGTCTTGGAAGTTAGAAC ACACTCCATGGGTGACGTGGGATGTAGCTCATTGGGGGTCCTTTACCCCATTTTTGTGTCAGTCTATACC CTACACTAGTGAAAGACAAAACAAAAAAATATTGAAATTGGCTCAGTCTCTCATGGGAATCCCAAGATGG GTTTAGCCTTTTGGACCTACTCTAATATGCCCAAATCAAGAATTATCTTAGAAGTACAAGGATCCATGCC AACTTGCCTTTTAGTGGATTTCTTGGCTTTCTTGAAAGTATCCTCTAAAAAACATTTTTAAATGTTTGGG CGTTTAAAAATTATTTATGTGTGGTGTGAAGAGTAGGCAGCACCTATCTTAAGAATTCTGTGTGACCAGG AAAGGAGGTGGAATTCCTGAACAGAAGCAATGCAAATACACAGAGAACTTGGTGAGTTGGGGTCATATGT GGATATGTAAATGGTCTATGCTGTATAACTTATTACAGGCATGAGCGATTGTATGCCTGTGTTAACTCTG CTGTGTCAAAGTCATTTCCTATTTAAACTGAATTTGTAGACCACGGAAGAGGATCAGGATTAATTATGTC TGTGAGGAAGAAGAAAAGCAAATAGCTCACTTGATTAAATACATTACAATAATCACTACATTCTGTCCTT TAGACCTGTGCTGTCCAATAGGGTAGCCACCAGACCTTATGGTCACTGAGCACCTAAAATGTGCTCCTAG TACAACTGAGAGACTGAATTTTAAATTTTACTTAATTTTTTAACAGTTAAATTTAAATTTAAAAGCTGAT TCTCAGCTCAATTATTGGAAAATGTTAAGTACGCTTGGAATAATTTGGGTATATACACCTATTTTTTTCA ATTATGAATTTTATGACATCTAAATATAGATCACCTATTTCTGGTAAAATTTAGCATCTGAATTGAGATA TATTGTAGGTATAAAATACACACTGGACTTCAAAGACTTAGTATAAAAAAGGAATGTAAAATATCTCAAT AATGTTTATATTGATGATGTGTCAAAATGATAATCTTTTGTATGTATTGGGTTAAATAAAACATATTATT AAAATGAATTATACCTGCTTCTTTTTACTTTTTTAATGTGGCTATAGAAAATTTTAAATTACAATTACGG TTCATATACTGTTTCTATTGACAGCACAGCCCTAGACTAAGCCGCTTTGGAATTCAAGAGGGAGTAAGTG ATGAGGTTCCATTGACTGGGTAGTCTTCAAGTGTGCATGGTGCTAACTCTCTGAGCCCAAAGTGAATGAG TGGTCAGTCAGGTTCCAGGCAGGTTATAGGGCAGGATGCAACGCCGGGTAAGGTCATTCTGCACATGTTA ATTGTTGATGTAGAACTCTTTCCTGCGTGGGTAGGGCCCTTGTACAGCAAAAGCAGCTGAAGTTTGTCTT TCAGGGACCTGAGGGGTAATTCATTTAATTGTGACTGTAAACTGAAATGGCTAGTGGAATGGCTTGGCCA CACCAATGCAACTGTTGAAGACATCTACTGCGAAGGCCCCCCAGAATACAAGAAGCGCAAAATCAATAGT CTCTCCTCGAAGGATTTTGATTGCATCATTACAGGTAATGTACTCATCATCATTCCACCTCAAAAAATTA AAATAAGGGCTACCTTTTTTTTTCATGTGCAGGAACTGATATTTTTTATATCTTAAAAACTAGCTGAGCA ACTTAATTTTAAAAAAATTATGAACCATTGACAATGCTTCATGTGTTTAAAAGTAAAATGGCCATTTTAC TATTCTCTGAAATAAATGTATTTCTTGTGAATATTTAAGATCTAGCCAAGGAAAGAGGTATTAGCTCACA GTTACTTATTATTTCCTATTTTTGCAGAATTTGCAAAGTCTCAAGACCTGCCTTATCAATCATTGTCCAT AGACACTTTTTCTTATTTGAATGATGAGTATGTAGTCATCGCTCAGCCTTTTACTGGAAAATGCATTTTC CTTGAATGGGACCATGTGGAAAAGACCTTCCGGAATTATGACAACATTACAGGTATGAAAAGCCTAATGA TATTTTGAGTGGTAATTTAAACTGCTTCAGCCAAGGGCAACAAGGAAAGATGAGTGGTATAGGGGAATGC TTTAGCATTTGAATTCCAACTCTACCATTTGTTAAGTGACCTGGGATTGTTCTCTAAGCCATAATTTCCC CACTTGTAAAACGGGAATAATATTAATACTTAACTCACAAGGTTGTTAATAAGATCATGTGAACACATAT TCAAGAAACTTAGCATAGGGTCTAGTATAATACATAAACAGTAAATATTTGATTATTTTTCTTCAAATAA AAGTATTGGATGGTGAATCATGAGTTCTAGTCCAGGTTCTGCCACAGTCTATTTGGGCAAGCTGCTCATA CTCACTGACACCCCTCAACCCCTTTTTCTTAACTATAAAATAAGAAGGTTATTTTAGATCAATAAGTCTC AAATTTGTTACACAAACGTGTCTTCTACAAAATGTTAATGTATGCCTTGTAATGAAAAGGATTTTGTGGT CCAATAAATTCATGAACCATATCTTGGTCTTGTAGGTTCATAATACAGACATCTGTGGATCTGAGAAGTC CTGCAGTGAGGAAATTTTTCCAATATTTTTCACCCTTAATTTGGCCATGGAATTTAATTCTTTCTTTTCT TTTCTTTTTTCTTTTTTTTTCTTTTTTTTTTTTTTTTTGAGACAGAGTCTCGCTCTGTCACCCAGGCTGG AGTGCAGTGGCACAATCTCAGCTCACTGCAACCTCCAACTCCCGGGTTCAAGCAATTCTTCTGCCTCAGC CTCCCAAGCAGCTGGGACTACAGGTGCGTGCCACCATGTCCAGCTAATTTTTGTATTTTTAGTACAGACA GGGTTTCACCATATTGGCAAAGCTGGTATTGAACTCCTGACCTCATGATCCGCCCACCTTGGCCTCTCAA AGTGCTGCCATTACAAGCGTGAGCCACCGCACCCGGCCGGAATCTAATTCTTTCTATGAAAACTTTTACT TATCTGGATCTCTCTTTTTTTTTTTTTTTTTTTGAGATGGAGTCTTGCTCTGTTACCCAGGCTGGAGTGC AGTGGCGTGATCTGGGGTCAATGCAACCTCAGCCTCCTGGGTTCAAGCGATTCTTGTCCTTCAGCCTCCC AAGTAGCTGGGACTACAGGCATGTGCCACCACGCCCTAATAATTTTTTGTATTTTTAGTAGAGATGGGGT TTCACCAAGTTGGTCAGGCTGGTTTCGAATTCCTGGCCTCAAGTGATCCACCCGCCTTGGCCTCCCAAAG TTCTGGGATTACAAGAGTAAACCACCTTGCCTGGGCTTACTTAGCTGATCTCTTAAGTTTCTTTCCCATT TTAATAATTTTTAAAATTATAAGTCATAAGTTAGAACATAGGGCTCCCTCTACAATTTTTAATATCAGTT TTAGCTTCTAATACCTTTAAATCTGCTCTCAAGAGCACTCACACCTCCCCATAAAAGGTAACCCTTTAGA CAAGGCATGGAGGGTCATGATAGATTTTGCTGGTAGCCATCACCCTCATTGCTCAAAGGACCAAGACAGG TCTGGGGCTTGCTTGAGGCTGGGGTTGCTTAAATAAAGGGATCCCTTCCAAACTGAGACCCTTTTGGAAT CCTCCTTTCTTTTAGAATCCTCCTCTCTGAAGGAAGAAGGTCATGCTTCCTTAGGCCCCCATAAGTTTTC CAGTGGACTATAGCAGGGTTTTGTGCGGTCGAAACCAGGCTTTTCTTACTTGTGTATGGTTCATCAATTT CTTTGAATGACTCTTTAGATGAGCACTTACAAGAGAAATAAATCATACATCTGCAAACTTTCTGAGGGCA TCTCAGAGCTCATCTTGTCTAGGATGATGTCTTAGTCCCTTCCTGCTGCTGTAACAAAAAATACCTTGAC

TGCGTAATTTATAAGAACAGAAATTTATTTCTCATAGTTCTGGAGGCTGGGAACTCCATGATCAAGGCAC CAGGAGATTTAGGGTCTGGTGAGGGCTCACTCTCTGCTTCAAAGACGGTGCCTTCTCTGTGTCCTAACAT GGCAGAAGGGCAAAAAAAGATCTAACAGACTCGCTCAAGCCCTTTTGCAAGGGCACAAATTCCATTCATG AGTGTGGAGTCCTCATGATCTAATCACCTCCTAGACACCCCACTTCTTAATACTATTACACTGGGGATTA AGCTACAACATAAATTTTGGAGGGACAGAAACATTCCAACCATAGCAGATGACAAATATATGCCCCATAC TCAGTCATTATCCCTTCCTTCACTCTAGACAGGCACCACTAATCTATCATGGGAATCTTTCATGTAGAGT GCAGACACGGCCACAGAATCCTTCTCAACACAGTTCCCCAGACAATCACACGCAATGAGCCAATCAGTAT TGGTACATGTGTTGGAATCTGGTGGCCATCCTTAATCTAGTCTAAAGTCTCATTGTACAGAAAAGAAAAC TGAGCCTCAGAGAAGTTAAATGATTTTCTTAAGATCATACAATCAGTTTAGCCCTGAAAAGAGTCAGAAA TAAAACTATTTTGCCTATGGATGAAGCCAATTAGTAGAAAGAATTAATGTAGAACTTTCTGCCCCGCACT AGCGCTGTAGTCAGAAGAGGCCTAAAGCCCAGATTTGTTCAGCTGTCTCTGGATTTTCTAGAGCCAAACT CCTGCAAGATCAGAATCTACTGATAAATGTCAAACTCTTCCCCAAAGGATACCAAGCTAGGCCACTGACA GATCCCACATCATTTTCTGTGATTTTCCAAAATAACTTTCAGGAGCAGTGCCCATGGCTGCACCCTTCTT TCGATTTTTTTAAAGATTCCTTTTCCTTCTCTTCACTCCTGCCACCAGTGACCTAAAGCTCCAGAAAAGT GACAAGTGGCATCTCCTTTCTTTGCAGGGGAAAGGCAAATTATTCTAGAGCAGGAAGCAGTTCAGACTTA GAGCAAAACAATTACAGTAGAGCCATGGGCTACCCAGACCACAACGCTAGCCTTTGGTTATCTAAGATCT TAGATAAACCAAATTTGAAGTCCTGGAATAGCTAACTTGTCCCCAGCTTGGGTTAAAAGCATAGAGATGC TGGTAGGCCTGGCTTTGAATCCCCAACTCTGCCTTCTGCTACCAGAACAGCCTCTCTGAGCTTCAGTTTA CTCATGTGGAAAAAGGATAATATTTATCTCTTAAGATTGTTGGGAAAATTAATGAGGTAAATTGAGAAAG CACATAATAGATGCTCAGTAAACAGTAGTTTTCATTAGAATTATGATTAACTCATTTGTGTCAGGGAAAC TAATGCTTTCTTTCTGCAAGAAGTAAGAGGAGAACACTCGGGCCATAGCTGTGTGTCCTGAACGTCTAGC GCAGGCTAGATATGACATTAACAACTCAATTGTTGGCAACAAGGCCCCACGAGGCACAAAGGCACGGAAC TTTAGGACACTGCTGCCATTGGGCTTTGGAGACCAGCCTTGCCTAATCAACCAAGGAGATCTCTTGTTTC AGGCTGATTTGGGTGGAAGTTGTATGGCCCCAGCCCGCATGTTTACATGCTCCAAAAGAGGATGGCCACA CAACTAATCTCTCCTTTGTCTTTTCCCAGGCACATCCACTGTAGTATGCAAGCCTATAGTCATTGAAACT CAGCTCTATGTTATTGTGGCCCAGCTGTTTGGTGGCTCTCACATCTATAAGCGAGACAGTTTTGCAAATA AATTCATAAAAATCCAGGATATTGAAATTCTCAAAATCCGAAAACCCAATGACATTGAAACATTCAAGAT TGAAAACAACTGGTACTTTGTTGTTGCTGACAGTTCAAAAGCTGGTTTTACTACCATTTACAAATGGAAC GGAAACGGATTCTACTCCCATCAATCCTTACACGCGTGGTACAGGGACACTGATGTGGAATATCTAGAAA TAGTCAGAACACCTCAGACACTCAGAACGCCTCATTTAATTCTGTCTAGTAGTTCCCAGCGTCCTGTAAT TTATCAGTGGAACAAAGCAACACAATTATTCACTAACCAAACTGACATTCCTAACATGGAGGATGTGTAC GCAGTGAAGCACTTCTCAGTGAAAGGGGACGTGTACATTTGCTTGACAAGATTCATTGGTGATTCCAAAG TCATGAAATGGGGAGGCTCCTCGTTCCAGGATATTCAGAGGATGCCATCGCGAGGATCCATGGTGTTCCA GCCTCTTCAAATAAATAATTACCAATATGCAATTCTTGGAAGTGATTACTCCTTTACTCAAGTGTATAAC TGGGATGCAGAGAAAGCCAAATTTGTGAAATTTCAGGAATTAAATGTTCAGGCACCAAGATCATTCACAC ATGTGTCCATTAATAAGCGTAATTTTCTTTTTGCTTCCAGTTTTAAGGGAAATACACAGATTTACAAACA TGTCATAGTTGACTTAAGCGCATGAGACACCAAATTCTGTGGCTGCCATCAGAAATTTTCTACAGTACAT GACCCGGATGAACTCAATGCATGATGACTCTTCTTATCACACTTGCAAATGAATGCCTTTCAAACATTGA GACTGCTAGAACCAAGCACTACCAGTATCTCCATCCTTAACTGTCCAGTCCAGTGATGTGGGAAGTTACC TTTTATAAGACAAAATTTAATTGTGTAACTGTTCTTTGCAGTGAAGATGTGTAAATAAGCGTTTAATGGT ATCTGTTACTCCAAAAAGAAATATTAATATGTACTTTTCCATTTATTTATTCATGTGTACAGAAACAACT GCCAAATAAAATGTTTACATTTTCTTTCATA

La secuencia del mRNA es la siguiente,

>gi|171846229|ref|NM_005097.2| Homo sapiens leucine-rich, glioma inactivated 1 (LGI1), mRNA GAAAGCAGAAAGCTGTTTATTTTCTGTGACATCACTTCACTTTGCCTTCGAAGGCTGGTCTGTGCTCAGT GTTTTCGTGGTGATGCAAGTCGGCTCTCTCCTCCAGCAGTTGGATCCCTCCCATCTCACAGTACCTCACA GGTCTCTTCCCCCGAGCAGTGCATTGCTGGAGCGAGGAGAAGCTCACGAATCAGCTGCAGGTCTCTGTTT TGAAAAAGCAGAGATACAGAGGCAGAGGAAAAGGGTGGACTCCTATGTGACCTGTTCTTAGAGCAAGACA ATCACCATCTGAATTCCAGAAGCCCTGTTCATGGTTGGGGATATTTTCTCGACTGCATGGAATCAGAAAG AAGCAAAAGGATGGGAAATGCCTGCATTCCCCTGAAAAGAATTGCTTATTTCCTATGTCTCTTATCTGCG CTTTTGCTGACTGAGGGGAAGAAACCAGCGAAGCCAAAATGCCCTGCCGTGTGTACTTGTACCAAAGATA ATGCTTTATGTGAGAATGCCAGATCCATTCCACGCACCGTTCCTCCTGATGTTATCTCATTATCCTTTGT GAGATCTGGTTTTACTGAAATCTCAGAAGGGAGTTTTTTATTCACGCCATCGCTGCAGCTCTTGTTATTC ACATCGAACTCCTTTGATGTGATCAGTGATGATGCTTTTATTGGTCTTCCACATCTAGAGTATTTATTCA TAGAAAACAACAACATCAAGTCAATTTCAAGACATACTTTCCGGGGACTAAAGTCATTAATTCACTTGAG CCTTGCAAACAACAATCTCCAGACACTCCCAAAAGATATTTTCAAAGGCCTGGATTCTTTAACAAATGTG GACCTGAGGGGTAATTCATTTAATTGTGACTGTAAACTGAAATGGCTAGTGGAATGGCTTGGCCACACCA ATGCAACTGTTGAAGACATCTACTGCGAAGGCCCCCCAGAATACAAGAAGCGCAAAATCAATAGTCTCTC CTCGAAGGATTTCGATTGCATCATTACAGAATTTGCAAAGTCTCAAGACCTGCCTTATCAATCATTGTCC ATAGACACTTTTTCTTATTTGAATGATGAGTATGTAGTCATCGCTCAGCCTTTTACTGGAAAATGCATTT TCCTTGAATGGGACCATGTGGAAAAGACCTTCCGGAATTATGACAACATTACAGGCACATCCACTGTAGT ATGCAAGCCTATAGTCATTGAAACTCAGCTCTATGTTATTGTGGCCCAGCTGTTTGGTGGCTCTCACATC TATAAGCGAGACAGTTTTGCAAATAAATTCATAAAAATCCAGGATATTGAAATTCTCAAAATCCGAAAAC CCAATGACATTGAAACATTCAAGATTGAAAACAACTGGTACTTTGTTGTTGCTGACAGTTCAAAAGCTGG TTTTACTACCATTTACAAATGGAACGGAAACGGATTCTACTCCCATCAATCCTTACACGCGTGGTACAGG GACACTGATGTGGAATATCTAGAAATAGTCAGAACACCTCAGACACTCAGAACGCCTCATTTAATTCTGT CTAGTAGTTCCCAGCGTCCTGTAATTTATCAGTGGAACAAAGCAACACAATTATTCACTAACCAAACTGA CATTCCTAACATGGAGGATGTGTACGCAGTGAAGCACTTCTCAGTGAAAGGGGACGTGTACATTTGCTTG ACAAGATTCATTGGTGATTCCAAAGTCATGAAATGGGGAGGCTCCTCGTTCCAGGATATTCAGAGGATGC CATCGCGAGGATCCATGGTGTTCCAGCCTCTTCAAATAAATAATTACCAATATGCAATTCTTGGAAGTGA TTACTCCTTTACTCAAGTGTATAACTGGGATGCAGAGAAAGCCAAATTTGTGAAATTTCAGGAATTAAAT GTTCAGGCACCAAGATCATTCACACATGTGTCCATTAATAAGCGTAATTTTCTTTTTGCTTCCAGTTTTA AGGGAAATACACAGATTTACAAACATGTCATAGTTGACTTAAGCGCATGAGACACCAAATTCTGTGGCTG CCATCAGAAATTTTCTACAGTACATGACCCGGATGAACTCAATGCATGATGACTCTTCTTATCACACTTG CAAATGAATGCCTTTCAAACATTGAGACTGCTAGAACCAAGCACTACCAGTATCTCCATCCTTAACTGTC CAGTCCAGTGATGTGGGAAGTTACCTTTTATAAGACAAAATTTAATTGTGTAACTGTTCTTTGCAGTGAA GATGTGTAAATAAGCGTTTAATGGTATCTGTTACTCCAAAAAGAAATATTAATATGTACTTTTCCATTTA TTTATTCATGTGTACAGAAACAACTGCCAAATAAAATGTTTACATTTTCTTTCATA

La siguiente informacin presenta la regin correspondiente al exn, la secuencia codigicante (CDS) y la protena traducida.

Mediante la herramienta clustalw se realiza un alineamiento entre la secuencia total del gen y la secuencia del ARNm.

Los resultados muestran un score de 100 entre la secuencia A (ADN) y la secuencia B (mRNA) , lo cual confirma que el mARN de 2366 pb es la transcripcin del gen ILGI1 de 47351 pb.

El alineamiento en detalle se presenta a continuacin,

CLUSTAL 2.1 multiple sequence alignment EpilpesyDNA 36000 EiplepsymRNA EpilpesyDNA 36060 EiplepsymRNA EpilpesyDNA 36120 EiplepsymRNA EpilpesyDNA 36180 EiplepsymRNA EpilpesyDNA 36240 EiplepsymRNA EpilpesyDNA 36300 EiplepsymRNA TTCTAAAGTTCATGAATGTTTGCACTTGAGAAACCTTATATTTAATTAGTGGAGGTAGAA ---------------------------------------------------------GAA 3 *** GACATAGACTTTTGAGAATGTTTTAAAGGGAAGTTGAGAGAATTTTCAAGGGTTTAGAGG AGCA------------------------GAAAGCTG------------------------ 15 ** * *** ** CTAATGGTATGGGCTCTTTAGTTTTAGTTTAGTTTTTTATTTTTATTGTGGAAAAAAAAA -----------------------------------TTTATTTT--CTGTG---------- 28 ******** **** ACACCTCTAAGAAAAGTTTATAACGTGTATATATATGGTGCTGGTTAAAAATAGAAAATT -----------------------------------------------------------ACACTACTTGGTGACCTTCCCCAGTCATACAGCATTAGCACTGTGCTATGTCTATATTTG ACATCACTT-----------------------------CACTTTGC-------------- 45 *** **** **** *** TGTACTAGGGACACATTTCCCTTTCCTTGACATTATCAGGACAATTTTTTACTCATTAAC ------------------------CTTCGA-------AGG-------------------- 54 * * ** ***

EpilpesyDNA 36360 EiplepsymRNA EpilpesyDNA 36420 EiplepsymRNA EpilpesyDNA 36480 EiplepsymRNA EpilpesyDNA 36540 EiplepsymRNA EpilpesyDNA 36600 EiplepsymRNA 101 EpilpesyDNA 36660 EiplepsymRNA 111 EpilpesyDNA 36720 EiplepsymRNA 141 EpilpesyDNA 36780 EiplepsymRNA 158 EpilpesyDNA 36840 EiplepsymRNA 176 EpilpesyDNA 36900 EiplepsymRNA 191 EpilpesyDNA 36960 EiplepsymRNA 198

CTTTATTAGAATCATTCTAGTGATTTACGGAAGAACAAATAGAAAGGGAAAGGGAAAAAC ----------------CTGGT--------------------------------------- 59 ** ** ATTCCTGTGATCACAGGAAAAGAACCAGAAGAAGGAGAAAGAGGGGGATGATTAAAGAGA ----CTGTGCTCA----------------------------------GTG---------- 71 ***** *** ** ATAGATGGATAGGTTTTTATAAGAAAACTTTTTTCTGAGGTGGAATGAATGGTGCTAGAA ------------------------------TTTTC----GTGG------TGATGCAAG-- 89 ***** **** ** *** ** AGGAAAAGGAGGCGTGGGCTTTAGAAAGGAGAAAGGATGTACTGGGGAAGAGATAGAGCT --------------TCGGCT---------------------------------------- 95 * **** ACTGGCACTGTGGGTTGGAGGTTTGGTTGCTCTCCGGAGCAAGCGAGGAGGAAGCAGGCA -----------------------------CTCTCC------------------------****** GAAGGGGCAGCCCAGGCAAGAGGAGGGGGCGCATTTTGGAGATGGACACTCACCAGTTTG ----------TCCAG--------------------------------------CAGTT-**** *****

GTTAGTCATCATTTGGGAGATATCCCCTAGATTGAGCGTCTGAGAGTGTGAATCTCTCGT -----------------GGATCCCTCCCA---------TCTCACAGT----ACCTCACAG *** * ** * *** * *** * *** *

GTTCTGTTCTAAATGGAAAGATCAATGCCCAGGAATCATTGTTGTTTTTAACCAACATAA GT-CTCTTCC-----------------CCCGAGCA------------------------** ** *** *** * *

AGTGTTGTTTAAATATTGAGTGCACCTGTAATCCCAGCACTTTGGGAGGCCGAGGCGGGT -------------------GTGCATT----------------------GCTGGAGCGAG***** ** * *** *

GGATCACTGAAGCTCAGGAGTTCAAGACCAGCCTGGCCAACATGGTGAAACCCTGTCTCT -GA-----GAAGCTCACGAAT--------------------------------------** ******** ** *

ACTAAAAATACAACAATTAGCCGGGTATGGTGGCGGGCACATGTAATCCCAGCTACGGCA -------------------------------------------------CAGCTGC---***** *

EpilpesyDNA 37020 EiplepsymRNA 205 EpilpesyDNA 37080 EiplepsymRNA 221 EpilpesyDNA 37140 EiplepsymRNA 235 EpilpesyDNA 37200 EiplepsymRNA 251 EpilpesyDNA 37260 EiplepsymRNA 258 EpilpesyDNA 37320 EiplepsymRNA EpilpesyDNA 37380 EiplepsymRNA 278 EpilpesyDNA 37440 EiplepsymRNA 287 EpilpesyDNA 37500 EiplepsymRNA 299 EpilpesyDNA 37560 EiplepsymRNA 322 EpilpesyDNA 37620 EiplepsymRNA 339

GGCTAAGGCAGGAAAATCACTTGAACCCGGGAGGCAGAGGTTTCAGTGAGCTGAGGTCAC -------------------------------------AGGTCTC---------------**** ** ACCACTTCACTCCAGCCTGGGTGACAGAGCGAGACTTTTGTCTCAAGAAATAAAAGTAAA --------------------------------------TGTTTTGA-----AAAAGCAG*** * * ***** *

TAAATATGGAGTGGGTGTTGAAGTGAACAGGTTAGGAACTGAAAGGACAATAGCAGGAAT -AGATACAGAG-----------------------------------------GCAG---* *** *** ****

CTGAGTAGTAGTTCAGAAGAAAAAAAAAGGACCACAAGTCTTTTAGTAGGCTGGAAATGA ---------------------AGGAAAAGG---------------GTGGACT-------* ****** ** * **

CAAAAGAGACTCATCACTAAGCTTTATCATTAAATGCCTTTGTTTAATTTATCACTACAG ------------------------------------CCTATGT----------------*** *** TTTACATCACCTTTTTTTTTTTTTTTCCAGGAGCCTTGCAAACAACAATCTCCAGACACT -----------------------------------------------------------CCCAAAAGATATTTTCAAAGGCCTGGATTCTTTAACAAATGTGTAAGAGGACCTAAGAAA -------------------GACCTG--TTCTT-------------AGAG----CAAGA-* **** ***** **** ****

TCACTGAACAATTAATTAATTTGCAGTCATTAACAAGGAAGCCTGGTATTGATTATTATA -----------------------CAATCACCA---------------------------** *** *

ATTTTTAATTGGCTTTTAAATTAAAATTAGGATTTTTAAATCTAAATTTTAACTGGCCAA ----------------------------------------TCTGAATT--------CCAG *** **** ***

AAATTTAAATTTACATGGACTAACATTTGAGTCTTCTATGGTATTTATCATTGGGAATTC AAGCCC-----------------------------------TGTTCATGGTTGGGGAT-** * ** ** ***** **

TGAATACATGGCACCCAGGCATTCCTTTGACAAAGTGTGTGTCAATACGGCAGGGAGCGA --------------------ATTTTCTCGAC----TGCATG-------------------

*** EpilpesyDNA 37680 EiplepsymRNA 348 EpilpesyDNA 37740 EiplepsymRNA 368 EpilpesyDNA 37800 EiplepsymRNA EpilpesyDNA 37860 EiplepsymRNA 393 EpilpesyDNA 37920 EiplepsymRNA 417 EpilpesyDNA 37980 EiplepsymRNA 442 EpilpesyDNA 38040 EiplepsymRNA 455 EpilpesyDNA 38100 EiplepsymRNA 472 EpilpesyDNA 38160 EiplepsymRNA EpilpesyDNA 38220 EiplepsymRNA 486 EpilpesyDNA 38280

* ***

**

**

TGATAAAGGACTTTGACGGGGTGGGAAGAGACCCGGCTAGAAATTTCAAAATCCCTTTTT ---------------------------------------GAA---TCAGAA--------*** *** **

TCTCATGAGGTCCAGGAAATAGGGCTACAAAGGAAAAACATAAAGGAGTCTGGGAAACTG ---------------------------------AGAAGCA-AAAGGA---TGGGAAA--* ** ** ****** *******

ACAGAATAAAGATGAAAGGTTACATCCTTGGGTTGGTCAATCAACAGAGAAATGGAACAG -----------------------------------------------------------GAAGTGGGGGAAGGGAGAAAAGTTTGCCTTTTTTCTCATTAGACTGAACAACAACTCTAA ------------------------TGCCTGCATTCCC------CTGAA-AAGAATT---***** *** * ***** ** ** *

GCTTAATCCCCAGGAATTTGGCTTAAGCCCCAGGAAGGGAATGAACTTAATTATTTTTGA GCTTATTTCCTA------TGTCTC--------------------------TTATCT---***** * ** * ** ** **** *

CCCTTTTTCAAACAAAAGAGCTTTCAAACTTTGACTTTTAAGCCTCCGAGTGAAAGATCT -----------------GCGCTTTTG----CTGACT-----------GAGGGGAAGA--* ***** ***** *** * ****

TTGCAAGGAATTTGTATATTTTGCTGTAAGCAAAGCAAGTAATATATGCCTAAACTTCTC --------AACC----------------AGCGAAGC-----------------------C ** *** **** *

AAAAGCAATTCAGCTTCATTATTTGCCTTCCAGAACATTAGAAAGGTATTTAAATGTCAT AAAA-------------------TGCCCTGCCG---------------------TGT--**** **** * * * ***

AAGGTGGAAGACAGGAACAAATGAAAGGATATATGAAAACAAAAGCTTTTTTAAAAAATT -----------------------------------------------------------ATTCAGGTACTTGTCTGTAGAATATATATTCAAAACAACTTTTCTCCAATAACAAACAAA ------GTACTTGT----------------------------------------ACCAAA ******** * ****

AAGAAAATCTGAGTCAAGTACTTTCTTCCAAGCCCAGTATTTAACTTCAAATCAGTAGCC

EiplepsymRNA 488 EpilpesyDNA 38340 EiplepsymRNA 501 EpilpesyDNA 38400 EiplepsymRNA 522 EpilpesyDNA 38460 EiplepsymRNA 546 EpilpesyDNA 38520 EiplepsymRNA 569 EpilpesyDNA 38580 EiplepsymRNA 575 EpilpesyDNA 38640 EiplepsymRNA 581 EpilpesyDNA 38700 EiplepsymRNA 599 EpilpesyDNA 38760 EiplepsymRNA 619 EpilpesyDNA 38820 EiplepsymRNA 624 EpilpesyDNA 38880 EiplepsymRNA 635

GA---------------------------------------------------------* TAATGAAAAACTTTCTTTGTTCCTTGCAGTATTTCCCCAGCCCCTCCTCCATGTCTATTA TAATG-----------------------------------------CTTTATGT-----***** ** ****

CCTCAATTCCTGAATGATATGGTTTAATTCCAGGAGAATGCCCAAATCAGGATTCTTCCA ---------------------------------GAGAATGCCAGATCCA------TTCCA ********* * ** *****

AGAGAGTCCTGGACAGACCATAGTTATCCTCTCTGGTTTACAGTAGCCTGAAAGGTTGCC -------------CGCACCGT-----TCCTC-CTGAT-----------------GTTATC * *** * ***** *** * *** *

ACTCATTAGTGTGGGTCAGAGTTCTCTTGGGAGACTAGGTCAGTTCTGGGAGTGTCTTGG --TCATTA--------------TCCTTTGTGAGA----------TCTGG----------****** ** *** **** *****

AAGTTAGAACACACTCCATGGGTGACGTGGGATGTAGCTCATTGGGGGTCCTTTACCCCA --------------------------------------------------TTTTAC---***** TTTTTGTGTCAGTCTATACCCTACACTAGTGAAAGACAAAACAAAAAAATATTGAAATTG ----------------------------------------------------TGAAAT-****** GCTCAGTCTCTCATGGGAATCCCAAGATGGGTTTAGCCTTTTGGACCTACTCTAATATGC -CTCAG------AAGGGAGTT-----------------TTTT-----------------***** * **** * ****

CCAAATCAAGAATTATCTTAGAAGTACAAGGATCCATGCCAACTTGCCTTTTAGTGGATT -------------------------------ATTCACGCCA--TCGC-------TGCAGC ** ** **** * ** ** *

TCTTGGCTTTCTTGAAAGTATCCTCTAAAAAACATTTTTAAATGTTTGGGCGTTTAAAAA TCTTG------------------------------------------------------***** TTATTTATGTGTGGTGTGAAGAGTAGGCAGCACCTATCTTAAGAATTCTGTGTGACCAGG TTATTCAC---------------------------ATC---------------------***** * ***

EpilpesyDNA 38940 EiplepsymRNA 649 EpilpesyDNA 39000 EiplepsymRNA 662 EpilpesyDNA 39060 EiplepsymRNA 673 EpilpesyDNA 39120 EiplepsymRNA 677 EpilpesyDNA 39180 EiplepsymRNA 681 EpilpesyDNA 39240 EiplepsymRNA 691 EpilpesyDNA 39300 EiplepsymRNA 707 EpilpesyDNA 39360 EiplepsymRNA 722 EpilpesyDNA 39420 EiplepsymRNA 744 EpilpesyDNA 39480 EiplepsymRNA 749 EpilpesyDNA 39540

AAAGGAGGTGGAATTCCTGAACAGAAGCAATGCAAATACACAGAGAACTTGGTGAGTTGG ----------GAACTCCT------------------------------TTGATG-----*** **** *** **

GGTCATATGTGGATATGTAAATGGTCTATGCTGTATAACTTATTACAGGCATGAGCGATT ---------------------TGATCAGTG--------------------ATGA-----** ** ** ****

GTATGCCTGTGTTAACTCTGCTGTGTCAAAGTCATTTCCTATTTAAACTGAATTTGTAGA ------------------TGCT-------------------TTTA--------TTG---**** **** ***

CCACGGAAGAGGATCAGGATTAATTATGTCTGTGAGGAAGAAGAAAAGCAAATAGCTCAC ---------------------------GTCT----------------------------**** TTGATTAAATACATTACAATAATCACTACATTCTGTCCTTTAGACCTGTGCTGTCCAATA -----------------------------------------------------TCCA--**** GGGTAGCCACCAGACCTTATGGTCACTGAGCACCTAAAATGTGCTCCTAGTACAACTGAG ------------------------------CATCTAGAGT-------------------** *** * * AGACTGAATTTTAAATTTTACTTAATTTTTTAACAGTTAAATTTAAATTTAAAAGCTGAT ------------------------ATTTATTCATAG--------------AAAA-----**** ** * ** ****

TCTCAGCTCAATTATTGGAAAATGTTAAGTACGCTTGGAATAATTTGGGTATATACACCT ---CAAC-------------AACATCAAGTC----------------------------** * ** * ****

ATTTTTTTCAATTATGAATTTTATGACATCTAAATATAGATCACCTATTTCTGGTAAAAT ----------------AATTTCAAGACAT-------------AC---TTTCCGG-----***** * ***** ** **** **

TTAGCATCTGAATTGAGATATATTGTAGGTATAAAATACACACTGGACTTCAAAGACTTA --------------------------------------------GGACT----------***** GTATAAAAAAGGAATGTAAAATATCTCAATAATGTTTATATTGATGATGTGTCAAAATGA

EiplepsymRNA 753 EpilpesyDNA 39600 EiplepsymRNA 760 EpilpesyDNA 39660 EiplepsymRNA 797 EpilpesyDNA 39720 EiplepsymRNA 808 EpilpesyDNA 39780 EiplepsymRNA 815 EpilpesyDNA 39840 EiplepsymRNA 825 EpilpesyDNA 39900 EiplepsymRNA 830 EpilpesyDNA 39960 EiplepsymRNA 865 EpilpesyDNA 40020 EiplepsymRNA 925 EpilpesyDNA 40080 EiplepsymRNA 985 EpilpesyDNA 40140 EiplepsymRNA 1009

-------AAAG------------------------------------------------**** TAATCTTTTGTATGTATTGGGTTAAATAAAACATATTATTAAAATGAATTATACCTGCTT -----------------------------------TCATTAA-----------------* ***** CTTTTTACTTTTTTAATGTGGCTATAGAAAATTTTAAATTACAATTACGGTTCATATACT ---TTCACTT----------------GAGCCTTGCAAACAACAATC----TCCAGACACT ** **** ** ** *** ***** * ** * ***

GTTTCTATTGACAGCACAGCCCTAGACTAAGCCGCTTTGGAATTCAAGAGGGAGTAAGTG ------------------------------------------CCCAAAAGATA------*** ** *

ATGAGGTTCCATTGACTGGGTAGTCTTCAAGTGTGCATGGTGCTAACTCTCTGAGCCCAA -----------------------TTTTCAA-----------------------------* ***** AGTGAATGAGTGGTCAGTCAGGTTCCAGGCAGGTTATAGGGCAGGATGCAACGCCGGGTA -------------------------------------AGGCCTGGAT------------*** * **** AGGTCATTCTGCACATGTTAATTGTTGATGTAGAACTCTTTCCTGCGTGGGTAGGGCCCT ------------------------------------TCTTT------------------***** TGTACAGCAAAAGCAGCTGAAGTTTGTCTTTCAGGGACCTGAGGGGTAATTCATTTAATT -----AACAAA-------------TGT-------GGACCTGAGGGGTAATTCATTTAATT * **** *** **************************

GTGACTGTAAACTGAAATGGCTAGTGGAATGGCTTGGCCACACCAATGCAACTGTTGAAG GTGACTGTAAACTGAAATGGCTAGTGGAATGGCTTGGCCACACCAATGCAACTGTTGAAG ************************************************************ ACATCTACTGCGAAGGCCCCCCAGAATACAAGAAGCGCAAAATCAATAGTCTCTCCTCGA ACATCTACTGCGAAGGCCCCCCAGAATACAAGAAGCGCAAAATCAATAGTCTCTCCTCGA ************************************************************ AGGATTTCGATTGCATCATTACAGGTAATGTACTCATCATCATTCCACCTCAAAAAATTA AGGATTTCGATTGCATCATTACAG-----------------------------------************************

EpilpesyDNA 40380 EiplepsymRNA 1012 EpilpesyDNA 40440 EiplepsymRNA 1072 EpilpesyDNA 40500 EiplepsymRNA 1132 EpilpesyDNA 40560 EiplepsymRNA 1175 EpilpesyDNA 40620 EiplepsymRNA EpilpesyDNA 44220 EiplepsymRNA 1235 EpilpesyDNA 44280 EiplepsymRNA 1295 EpilpesyDNA 44340 EiplepsymRNA 1355 EpilpesyDNA 44400 EiplepsymRNA 1415 EpilpesyDNA 44460 EiplepsymRNA 1475 EpilpesyDNA 44520 EiplepsymRNA 1535

TCTAGCCAAGGAAAGAGGTATTAGCTCACAGTTACTTATTATTTCCTATTTTTGCAGAAT ---------------------------------------------------------AAT *** TTGCAAAGTCTCAAGACCTGCCTTATCAATCATTGTCCATAGACACTTTTTCTTATTTGA TTGCAAAGTCTCAAGACCTGCCTTATCAATCATTGTCCATAGACACTTTTTCTTATTTGA ************************************************************ ATGATGAGTATGTAGTCATCGCTCAGCCTTTTACTGGAAAATGCATTTTCCTTGAATGGG ATGATGAGTATGTAGTCATCGCTCAGCCTTTTACTGGAAAATGCATTTTCCTTGAATGGG ************************************************************ ACCATGTGGAAAAGACCTTCCGGAATTATGACAACATTACAGGTATGAAAAGCCTAATGA ACCATGTGGAAAAGACCTTCCGGAATTATGACAACATTACAGG----------------******************************************* TATTTTGAGTGGTAATTTAAACTGCTTCAGCCAAGGGCAACAAGGAAAGATGAGTGGTAT -----------------------------------------------------------CACATCCACTGTAGTATGCAAGCCTATAGTCATTGAAACTCAGCTCTATGTTATTGTGGC CACATCCACTGTAGTATGCAAGCCTATAGTCATTGAAACTCAGCTCTATGTTATTGTGGC ************************************************************ CCAGCTGTTTGGTGGCTCTCACATCTATAAGCGAGACAGTTTTGCAAATAAATTCATAAA CCAGCTGTTTGGTGGCTCTCACATCTATAAGCGAGACAGTTTTGCAAATAAATTCATAAA ************************************************************ AATCCAGGATATTGAAATTCTCAAAATCCGAAAACCCAATGACATTGAAACATTCAAGAT AATCCAGGATATTGAAATTCTCAAAATCCGAAAACCCAATGACATTGAAACATTCAAGAT ************************************************************ TGAAAACAACTGGTACTTTGTTGTTGCTGACAGTTCAAAAGCTGGTTTTACTACCATTTA TGAAAACAACTGGTACTTTGTTGTTGCTGACAGTTCAAAAGCTGGTTTTACTACCATTTA ************************************************************ CAAATGGAACGGAAACGGATTCTACTCCCATCAATCCTTACACGCGTGGTACAGGGACAC CAAATGGAACGGAAACGGATTCTACTCCCATCAATCCTTACACGCGTGGTACAGGGACAC ************************************************************ TGATGTGGAATATCTAGAAATAGTCAGAACACCTCAGACACTCAGAACGCCTCATTTAAT TGATGTGGAATATCTAGAAATAGTCAGAACACCTCAGACACTCAGAACGCCTCATTTAAT

************************************************************ EpilpesyDNA 44580 EiplepsymRNA 1595 EpilpesyDNA 44640 EiplepsymRNA 1655 EpilpesyDNA 44700 EiplepsymRNA 1715 EpilpesyDNA 44760 EiplepsymRNA 1775 EpilpesyDNA 44820 EiplepsymRNA 1835 EpilpesyDNA 44880 EiplepsymRNA 1895 EpilpesyDNA 44940 EiplepsymRNA 1955 EpilpesyDNA 45000 EiplepsymRNA 2015 EpilpesyDNA 45060 EiplepsymRNA 2075 EpilpesyDNA 45120 EiplepsymRNA 2135 TCTGTCTAGTAGTTCCCAGCGTCCTGTAATTTATCAGTGGAACAAAGCAACACAATTATT TCTGTCTAGTAGTTCCCAGCGTCCTGTAATTTATCAGTGGAACAAAGCAACACAATTATT ************************************************************ CACTAACCAAACTGACATTCCTAACATGGAGGATGTGTACGCAGTGAAGCACTTCTCAGT CACTAACCAAACTGACATTCCTAACATGGAGGATGTGTACGCAGTGAAGCACTTCTCAGT ************************************************************ GAAAGGGGACGTGTACATTTGCTTGACAAGATTCATTGGTGATTCCAAAGTCATGAAATG GAAAGGGGACGTGTACATTTGCTTGACAAGATTCATTGGTGATTCCAAAGTCATGAAATG ************************************************************ GGGAGGCTCCTCGTTCCAGGATATTCAGAGGATGCCATCGCGAGGATCCATGGTGTTCCA GGGAGGCTCCTCGTTCCAGGATATTCAGAGGATGCCATCGCGAGGATCCATGGTGTTCCA ************************************************************ GCCTCTTCAAATAAATAATTACCAATATGCAATTCTTGGAAGTGATTACTCCTTTACTCA GCCTCTTCAAATAAATAATTACCAATATGCAATTCTTGGAAGTGATTACTCCTTTACTCA ************************************************************ AGTGTATAACTGGGATGCAGAGAAAGCCAAATTTGTGAAATTTCAGGAATTAAATGTTCA AGTGTATAACTGGGATGCAGAGAAAGCCAAATTTGTGAAATTTCAGGAATTAAATGTTCA ************************************************************ GGCACCAAGATCATTCACACATGTGTCCATTAATAAGCGTAATTTTCTTTTTGCTTCCAG GGCACCAAGATCATTCACACATGTGTCCATTAATAAGCGTAATTTTCTTTTTGCTTCCAG ************************************************************ TTTTAAGGGAAATACACAGATTTACAAACATGTCATAGTTGACTTAAGCGCATGAGACAC TTTTAAGGGAAATACACAGATTTACAAACATGTCATAGTTGACTTAAGCGCATGAGACAC ************************************************************ CAAATTCTGTGGCTGCCATCAGAAATTTTCTACAGTACATGACCCGGATGAACTCAATGC CAAATTCTGTGGCTGCCATCAGAAATTTTCTACAGTACATGACCCGGATGAACTCAATGC ************************************************************ ATGATGACTCTTCTTATCACACTTGCAAATGAATGCCTTTCAAACATTGAGACTGCTAGA ATGATGACTCTTCTTATCACACTTGCAAATGAATGCCTTTCAAACATTGAGACTGCTAGA ************************************************************

EpilpesyDNA 45180 EiplepsymRNA 2195 EpilpesyDNA 45240 EiplepsymRNA 2255 EpilpesyDNA 45300 EiplepsymRNA 2315 EpilpesyDNA 45360 EiplepsymRNA 2366 EpilpesyDNA 45420 EiplepsymRNA

ACCAAGCACTACCAGTATCTCCATCCTTAACTGTCCAGTCCAGTGATGTGGGAAGTTACC ACCAAGCACTACCAGTATCTCCATCCTTAACTGTCCAGTCCAGTGATGTGGGAAGTTACC ************************************************************ TTTTATAAGACAAAATTTAATTGTGTAACTGTTCTTTGCAGTGAAGATGTGTAAATAAGC TTTTATAAGACAAAATTTAATTGTGTAACTGTTCTTTGCAGTGAAGATGTGTAAATAAGC ************************************************************ GTTTAATGGTATCTGTTACTCCAAAAAGAAATATTAATATGTACTTTTCCATTTATTTAT GTTTAATGGTATCTGTTACTCCAAAAAGAAATATTAATATGTACTTTTCCATTTATTTAT ************************************************************ TCATGTGTACAGAAACAACTGCCAAATAAAATGTTTACATTTTCTTTCATATCCCAGAGG TCATGTGTACAGAAACAACTGCCAAATAAAATGTTTACATTTTCTTTCATA--------*************************************************** TAATTATTTACAGGAGCTGTTTCATTCTTGTTGTTGGTATGTTGAAGAAATAGTTTTATA ------------------------------------------------------------

Con la herramienta expasy tool se realiza una prediccin de la transcripcin con base al mRNA, debido a la naturaleza de cdigo gentico degenerado se presentan 6 frames, el frame elegido es aquel con menor nmero de stop codons.

La secuencia de aminocidos es corrida en Blast protein con el fin de identificar a que protena pertenece.

La protena con mxima identidad es NP_005088 Leucine-rich glioma-inactivated protein 1 coincidiendo con lo reportado en ncbi para el gen ILGI1.

Seguidamente se realiza un traduccin reversa partiendo de la proteina NP_005088 .1, se obtienen dos posible secuencias.

En la herramienta clustalw se realiza un alineamiento entre las dos secuencias obtenidas en la traduccin reversa y la secuencia de mRNA correspondiente a LGI1.

Entre la secuencia Reversa1 y la secuencia mRNA se logra el mayor score de alineamiento (74.0), seguidamente se corre en Blast-nucleotide la secuencia Reversa1 y se aprecia que evidentemente coincide con la secuencia del ARN mensajero del gen LGI1.

Con la herramienta expasy-tool calculamos el punto isoelctrico y el peso molecular de la protena.

Con la herramienta REBsites identificamos las enzimas de restriccin que menor nmero de fragmentos produce en la secuencia de DNA del gen LGI1

Para el diseo de primers se tiene en cuenta la regin del mRNA correspondiente a la secuencia codificante (CDS) que comprende las regiones entre los nucletidos 337 y 2010.

Los primers con mayor porcentaje de GC, es decir los ms estables son mostrados en la siguiente imagen.

Referencias [1] Mutant LGI1 inhibits seizure-induced trafficking of Kv4.2 potassium channels. Smith SE, Xu L, Kasten MR, Anderson MP. J Neurochem. 2012 Feb;120(4):611-21. doi: 10.1111/j.1471-4159.2011.07605.x. [2] Familial temporal lobe epilepsy with psychic auras associated with a novel LGI1 mutation.Striano P, Busolin G, Santulli L, Leonardi E, Coppola A, Vitiello L, Rigon L, Michelucci R, Tosatto SC, Striano S, Nobile C. Neurology. 2011 Mar 29;76(13):1173-6. [3] Electroclinical characterization of epileptic seizures in leucine-rich, glioma-inactivated 1-deficient mice. Chabrol E, Navarro V, Provenzano G, Cohen I, Dinocourt C, RivaudPchoux S, Fricker D, Baulac M, Miles R, Leguern E, Baulac S. Brain. 2010 Sep;133(9):274962. Epub 2010 Jul 21. [4] The leading edge of epilepsy research. Ikeda A. Rinsho Shinkeigaku. 2011 Nov;51(11):993-6. [5] Epilepsy gene LGI1 regulates postnatal developmental remodeling of retinogeniculate synapses.Zhou YD, Zhang D, Ozkaynak E, Wang X, Kasper EM, Leguern E, Baulac S, Anderson MP. J Neurosci. 2012 Jan 18;32(3):903-10. [6] Low penetrance and effect on protein secretion of LGI1 mutations causing autosomal dominant lateral temporal epilepsy. Di Bonaventura C, Operto FF, Busolin G, Egeo G, D'Aniello A, Vitello L, Smaniotto G, Furlan S, Diani E, Michelucci R, Giallonardo AT, Coppola G, Nobile C. Epilepsia. 2011 Jul;52(7):1258-64. doi: 10.1111/j.1528-1167.2011.03071.x. Epub 2011 Apr 19. [7] Autoimmune epilepsies. Irani SR, Bien CG, Lang B. Curr Opin Neurol. 2011 Apr;24(2):146-53. Review.

[8] The temporal and spatial expression pattern of the LGI1 epilepsy predisposition gene during mouse embryonic cranial development. Silva J, Wang G, Cowell JK. BMC Neurosci. 2011 May 13;12:43. [9] LGI2 truncation causes a remitting focal epilepsy in dogs. Seppl EH, Jokinen TS, Fukata M, Fukata Y, Webster MT, Karlsson EK, Kilpinen SK, Steffen F, Dietschi E, Leeb T, Eklund R, Zhao X, Rilstone JJ, Lindblad-Toh K, Minassian BA, Lohi H.PLoS Genet. 2011 Jul;7(7):e1002194. Epub 2011 Jul 28. [10] Similarity of molecular phenotype between known epilepsy gene LGI1 and disease candidate gene LGI2. Limviphuvadh V, Chua LL, Rahim RA, Eisenhaber F, Maurer-Stroh S, Adhikari S. BMC Biochem. 2010 Sep 24;11:39.