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Transcription maschine
MOLECULAR BIOLOGY
Part II
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Content
Translation
Mutation
DNA repair system
Manipulation on DNA and protein
Translation
COMPONENTS OF
TRANSLATION
TRANSLATION
mRNA
carries the genetic information transcribed
from DNA in the form of a series of codons.
codons.
2. tRNA.
Each AA has its own subset of tRNAs, which
bind the AA and carry it to the growing end of
a polypeptide chain if the next codon in the
mRNA calls for it. Each specific tRNA molecule
contains anticodon
anticodon,, that can basebase-pair with
its complementary codon in the mRNA
1.
Component of
Translation:
-mRNA
-tRNA
-Ribosom
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tRNA
- a hairpin structure
- amino site at (3 end)
- anticodon site, that bind with
codon in mRNA
There are 61 type tRNA in cell
3. rRNA
- associates with a set of protein to form
ribosomes - move along an mRNA molecule,
catalyze the assembly of AA into polypeptide
chains..
chains
- Ribosomes are composed of a large and
small subunit, each of which contains its own
rRNA molecules.
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Translation Proses
Initiation
Elongation
Termination
INITIATION
Binding of mRNA on
small subunit
ribosom
Binding of tRNA
inisiator at the P site
of ribosom
Binding of big
subunit ribosom
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ELONGATION
TERMINATION
- Codon
recognition
- Peptide bond
formation
- translocation
THE LAST STEP OF TRANSLATION
RIBOSOME (SITE A) REACH STOP CODON
(UAA UAG UGA)
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MUTATION
MUTATION,
Type of mutation:
1.Base pair (nucleotide pair) substitutions
2.Frameshift mutations
3.spontaneous mutation
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silent M,
M,
not resulting in a new aa (amino acid) in
the protein sequence,
sequence, eg. GCA
GCA or GCG
GCG.
2. missense mutation
mutation..
result in an aa substitution
substitution ex. CT
CTCCAC
in the DNA sense strand (glutamate
(glutamatevaline).
1.
3. a nonsense mutation
mutate an amino acid codon to a
termination codon or vice versa.
versa.
4. Mutation occurs in promoters or 5'
regulatory regions of genes or in introns
may affect their transcription, translation,
or splicing.
splicing. Ex.
Ex.beta
beta--thalassemias, result of nonnon-
2.Frameshift mutations
3.Spontaneous mutation
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cont
spontaneous base degradation.
degradation.
The deamination of cytosine to uracil.
spontaneous DNA damage via alkylation
the addition of alkyl (methyl, ethyl, occasionally
prophyl) groups to the bases or backbone of
DNA.
Alkylation can occur through reaction of
compounds such as SS-adenosyl methionine with
DNA. Alkylated bases may be subject to
spontaneous breakdown or mispairing
mispairing..
REPAIR" mechanisms :
Direct repair
Simple removal of damage.
Excision repair
Recognition by enzyme, followed by
excision. Glycosylases, uracil
glycosylase-glycosylase
--enzyme
enzyme which removes
uracil from DNA, AP endonuclease.
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plasmid
RESTRICTION ENDONUCLEASE
Type II restriction endonuclease
Palindromic sequences
Recognition site/cleavage
site/restriction site
Type of cut end
5
5--phosphat extention (Eco
(EcoRI)
RI)
3hydroxyl extention (Pst
(PstI)
I)
Blunt
Blunt--end (Pvu
(PvuII)
II)
Def:
Cyclic, selfreplicating
DNA, double
stranded,maintained
in the bacterial cell as
a extrachromosomal
DNA.
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Basic
priciples
of DNA
cloning
in
plasmid
vector
Human genomic
DNA lybrary
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