Tips for ARS mains answer writing

You are supposed to answer 66 qns in 3 hrs i.e. 66 qns in 180 minutes
40 X 2 = 80 marks obj- in 20-25 minutes
20 X 5 =100 marks obj-in-95-100 minutes
6 X 10= 60 marks obj-in- 60 minutes
- time management is very very imp.
In your answer use bullets alongwith diagrams, figures.
Where steps or process is asked go for flow charts & steps format
Where ever application are asked - use recent examples & Research achievements
& go for bullets
Enrich your answers with sub-points & examples.
underline your example & relevant or main step or information.
For subjective type answer like
Scope of transgenic in crop improvement - go for articles & magazines & try to
make note of given examples & arguments (for & against)
Do Your home work
- Go for recent research examples & happening in ICAR & CSIR system relevant
to your field. This will also help your in your interview.
- Make your own handy note book related to your information updates, examples,
research findings & miscellaneous topics you consider as important. Revise this at
last moments of the exam (i.e. 3 days before the exam 3 times)
- Please do not revise outside the exam centre. Current in that your will head will
hinder the retrieval of the information stored in your long term memory.
Solved Answer sets
Set I
Set II
Set III
Set IV
Set V
Set VI
Set VII
Previous year questions unsolved
Set I
1. Electron microscope was discovered by M.Knoll & E.Ruska
2. Hall life of P
32
is 14.5 days & C
14
is 5570 yrs.
Hall life of P
33
is 25.4 days & H
3
is 12.46 yrs.
3. Phase contrast microscope was discovered by ______________ & used to
study _________ without ________________.
4. Dark ground microscope invented by _____________ & useful in the study
of _______________.
5. In monocots vascular bundles are absent & Cork Cambium is absent in
stem.
6. In prokaryotic DNA replication RNA primer is about ___________
nucleotides where as in Eukaryotes it is of ___________ nucleotides.
7. Size of okazaki fragments in Prokaryotes is 1000 nucleotides.
8. Fork movement rate in prokaryoles in 1000 bp/sec where as in eukaryots is
100 bp/sec.
9. Example of negatively charged aminoacid is Glutanic acid & Aspartic acid
with single word symbol __________ & ___________ respectively.
10. Phynylatanine, tyrorine & tryptophan are the three aromatic amino acids.
11. Arachidonic acid has 18 no. of carbons with double bonds at 9,12 & 15.
12. Tryptophan is converted to Anxin which in turn contributes to formation of
__________ & ___________.
13. Sphingo sine is derived from Ceramide.
14. Leucine & Lyrine are purely ketogenic aminoacids.
15. Pericentric inversion involve centromere.
16. Tertiary trisomic condit
n
is (2n+1+1) (2n+2, 2n+1+1, 2n+1+1+1)
17. Chemical formula of Colchicine is ________________.
18. Quasidiploid is ________________ (2n+1-1, 2n-2, 2n-1+1)
19. Mendel did not work in _________ but worked in _______________ Pisium
Sativum & Lamar king.
20. Jumping gene hypothesis was given by __________ in ____________.
21. Binzor gave concept of recan muton & cistron.
22. 5 Bromo-uracil & O
6
methylguanive are the two example of base analoge.
23. Mutation was Ist discovered in Sheep (Encon) by seth wright & termed
cotned by Hug de Vries.
24. Meristem has high auxin content & have absence of Plasmodesmata.
25. For regeneration of Bt-cotton the variety used in tissue culture was Cocker.
26. Exposure of DNA to UV light leads to formation of Thymidine dimmers
(T=T).
27. Area under transgenic in India is 10.3 million lactave (5th rank)
28. Which one is more plymorphic SSR (SSR, SNP, AFLP, RAPD).
29. Most abundant marker is SNP (SSR, SNP, ALFP).
30. RAPD, ISSR uses single primer (True/False).
31. You may get poly morphosin without any variation in DNA deqnences
(True/False).
32. For determining copy number of a gene me can use Southern Blot & qPCR
techniques.
33. Define gene?
According to Singer and Berg in 1992. A gene is a combination of DNA sequence
that together constitute an expressible unit, whose expression leads to the
formation of one/more specific functional products that may be either RNA
molecule or polypeptide chain.
34. Define proteins & carbohydrates
Proteins are the polymers of amino acids linked by amide linkage [C
11o
-NH]
having N terminal (NH
3
) and C terminal (each) endo.
Carbohydrates are the hydrates of carbon or the compounds which give them upon
hydrolysin [CnH2] or polyhydroxy aldehyde/Kelones.
35. Av. Deight of an Amino acid is 110 Da
36. Cloning capacity of YAC, PAC, Cosmid, & PUC is 1000 Kb 300Kb 40Kb
& 4-5 Kb respectively.
37. YAC was developped by David Burke & BAC was developed by Melsimon.
38. Plitmus & PAT are examples of expression rectors.
39. Mito Chondria was discovered by __________ & is called Power house of
cell.
40. Enzymes act by lowering the activation energy of ryn.
5 Markers 20 Qns
1. How the cell cycle is regulated & controlled? Elucidate
Process of cell-growth and division is called cell-cycle. Cell phase involve - G1,
G2, S, and M phase.
Cell cycle is regulated various chekc points
mainly G1 - Check point
G2 - Check Point
M - Check Point
In G1 cdk levels are low, new pre RC complex not found and already found pre-
RC are activated for replication.
2. Write steps of Southern hybridization?
Destron sulphate used for enhancing hybridization.
Aried milk; hepanin; SOS prevent non-specific bending
3. How Enzymes catalyze reactions? What are allostearic Enzymes
Enzymes catalyze reactions by means of lowering the activation energy [Ea]
required to overcome the intermediate [ES] complex and form the product with out
being utilized in the ran.
Allosteric Enzymes: Enzymes which have an additional site called allosteric site
other than the acture site (bending to substrate) to catalyze the oxn.
Allosteric site (site for binding of effector molecules).
Activators (if molecule attachment enchances enzy. actinls)
Inhibitors (if reduces whibits)
Thereby effectors modulate the activity of allostrie enzymes eq. lac. repress of
lacoperon (allolactose inhisitor).
4. Compare & contrast between plant cell & prokaryotic cell?
Prokaryotic cell Plant Cell
At-cellular level:-
Nuclear material unbound as
naked.
cell-wall made of peptidoglycan.
organells absent
Mesosame as powerhouse 70s
Ribosomes
Genome Organization level
Intraw absent
No bound Histones
Genes are stacked and regulated
Bound as nucleus
Cell-wall made of cellular + hemi
cellulose
Present (eq. Milrechandius; er;
chloroplast etc).
Mitrchandria
80S Ribosames
Instrans present
Histones bound to proteins as
Nucleosomes
Censtacked genes.
by a common promotes eq. lac
operon
few retrotransposent/transposent
Insulators/Enhances absent
Functional level
Transcription and translation
simultaneously
Similarity
Heterotrophic except (Blue green
algaeycyanobactera)
Intergenic sequences and
retrotransposant, transposable
element at higher rate.
present
Separated by space. Transcription
in nucleus & translations in
cytoplasm.

Both perform the basic functions like replication; tran ripton, translations and
utilize energy in the form of ATP and are membrane bound. enclosed as cell.
6. On what levels the gene expression is regulated in Prokaryotes? Explain
Gene-expression in Prokaryolis is regulated at Transcription and Translation level.
At transcriptional level. Alongwith the RNA processing control initiation of
transcription of genes only when they are required depending upon the rate of
Metabolism, Response to environmental stress and cell-division.
7. Describe Lac openons give its mechanism of action? Explain with Diagram
Lac opiron is the Lactor metabolism system followed by Prokaryotes for caverting
lactose - glucox + galaclose (Energy sevrces)
(+ve) Regulation occurs when: [CAMP-CAP] binds to CAP site in Promoter and
activates/enhances adduct RNA Polymers to express lac Z, Y A glrus involved in
lactose metabolism.
Conditions
Glucose + ATP
CAMP
(-ve) Regulation Control
Lactox - Auolactole - Binds to lac Reprenor (Bound to promoler)
Conditions
Lactose present + Little amount of B-glutonidose
8. How will you generate a fine map rising marker for given 10 tns of the
chronisome?
Take divergent Parents 11x111
9. What is transposor tagging ? Explain
Determination of the region that is tagged by a Transposon in order words finding
the location of the region (DNA sequence) to which the transposon has been
integrated study of Inheritance and identification of the sequence tagged to
tranposon is important in Gene-isolation.
10. Write short notes on Paramutation?
Paramutation was the term used by R.A. Brink in 1950's.
Its involves the mutations that do not involve DNA sequence variation by variation
at histone modifications (Deacctylation/Methylation etc).
i.e. Justeraction b/w 2- alleles at a single locus, whereby one allile induces a
heritable change in other allile. The allile inducing the change is paramutagenic
while the allile that has been attered is called paramutant.
Since this change is meiotically inheritable, hence it violates mendels first low. A
paramutant allile may have an attered level of gene expression even though in the
segregating generation the paramutagenic allile is absent/gets separated from it.
Thus a single gene may result in spectrum of phenotypes. eq. r1 in maize from
completely colourlen to fully colored Kirquels.
In Maize paramutation seems to be RNA directed i.e. RNA directed DNa
mithylation carried by mop1 gene which is RNA dependent RNA polymerase
gene.
11. What is Chromosome walking chr jumping & chr. hoping? How they one
different
Chromosane walking: - When we have to find a gene out of the Candidate gems
and there is not much distance b/w the markus flanking the gene [Exonic region
mainly] then we head from the primers derived from market ends and reach to the
clone having both the marker end sequences same. The end of clone 1 derived
from one PCR act as primer sequence for other to generate clone2 and
subsequently.
Chromosane Jumping: - When we don't know where the gene is and probability of
large intrans/intergenic region or non-clonable gaps are there we go for chromosa
jumping
Genetic map is not saturated (condition) physical map not available.
Chromosome hoping:- When through chromosome jumping even we are not able
to reach to the gene, we go fs chromosome hoping where using translocation
stockes we hope into the gene, using sparse marker information.
Condition Sparse marker information
physical map unavailable.
12. What kind of Problems YACs suffer with? Give advantage of BACs over
YACs.
YaCs - Years artificial chromosames (David Burke in 1997) an imp cloning
cloning capacity - 1000Kb - 2000Kb eg. PYAC. Vector
TEL; CEN; ARS - Atonomously replicating sequences.
Problems
Chimeras are fsined [2 fragments from different regions.
Unitable and often incorporate in same.
Lead to deletions of Regions YAC.
Recombination is poorly understood thus YAC may integrate with yeast carasame.
Transformation is difficult
Maintainance is a problem.
Slow growing
Advantages of BAC's over YAC's
Easy to maintain and mainpulate
Chimiras not formed
CEN/TEL region not required
No delitions cloning capacity (250-300 Kb)
BAC- librarin created for sequening.
13. Describe process of replication in prokaryotes? How it is different from
Eukaryotes.
In Eukaryotes organisms there is replication origin as ARS intead of Oris in
Prokeyota.
No of Replicating regions in Enk. is many and in bidirectional bliay unlilee prok.
which show significance with Bidirectional movement.
Replication is initiated with DNA poly I with cere (L,E,Q) while in Eule, which is
(L, Q, E) which forms the core.
L, B, R, S) while DNA poly I, II, and (PCNA) as clamp loaden while Bclamp
Bsubmit.
14. What is translation? How it is regulated in Eukaryotes.
Translation:- Translation is the process in which cellular ribosanes create proteins.
It is an imp cap site part of gene expression i.e. mRNA produced by Binding trans
is decoded by a ribosame complex to produce a specific ammuni acid
chain/polypeptide which folds to fsm active protein.
Regulation of Translation in Eukahyotes:-
Initiation, Elongation and Termination factors
eIF4F process of scanning
eIF4F - eEF4E Binds to the 5'cap of mRNA
eEF4A - RNA helicase activity and removes secondary structure from mRNA
eEF4G Binds to eIF4E and PAB.
eIF4B - locates the first AUG site fs translation initiation
eIF3 - Ist factor bind to 4-S submit of Ribosame to facilitate subsequent steps
eIF2 - Facilitates binding of Met-tRNA met to 40S ribosamal subcent.
eIF5 - Promotes dissociation of all other IF's to carry out association 40S to 60S
eIF6 - Dissociation of inactive 80S ribosame into 60S & 40S
Elongation centrolled bf - eEFIL, eEF1B, r and eEFd
A codon on mRNA identifies a auti codar or trna (first genetic code)
15. Drawa fully featured structure of a Eukaryotic gene?


16. Elucidates different modes & methods of gene isolate? Give name & approches
of the method only.
Gene-isolation methods approach based. (Phentype to Genotype)
DDRT-PCR, SSH
Transcriptome sequencing
using next generation sequencing
454-Pysosequencing
Microarray (Hybridization based method)
SAGE - EST Clones (SAGE Tags)
Tlling;
forward genetics approach based (Genetype to phenotype)
MAP-based cloning
Insertional innitogeuesis
T-DNA nisestion
Transposen tagging
Retro transposan
Activation tagging
Positinal cloning based/Complementation tissuing
When gnome required we can use (PCR) and known primers to isolate gave.
17. What are molecular markers? What are its application in crop improvement.
Molecular markers are the differences in the nucliotide DNA at corresponding site
of the homologous chromosames which follow inheritance (mendelian) of patteus
(in case of nuclear DNA marker) and follow cytoplasmic/materul inheritance (in
case of organellar DNA markers).
Eq. SSR; RAPD; AFLP; SNP etc.
Applications of Molecular markers in trop improvement
Marker assisted selection:- Selection of plants with the desired gave at early stage
of growth can be done using inslecular marking date eq: MABC/MAR Sets
Selection of plants with heterozygons allelus at particular loci can be done.
Use of molecular markers in Map based cloning for isolation of gene eq. RFCP &
SSR.
DNA finger printing of Varieties for identification & characterization
Identification of cuttivars
Tagging of Gene using molecular markers. (Funcational marker eq. opaue 2 gene
using phiso)
Asessment of Genetic Diversity by random/specific markers. SSR marker
Gene introgression/QTL's DNA barcoding RAPD markers used
Detection and elimination of linkage drag. eq. finding donke recambinaty
Gene pyramiding Development of heterotic hybrids
eq. BLB and Blast re istance in Rice eq. Xa218X912 X95 genes.
18. What is PCR? What one its application
PCR is polymeose chain rxn (kavy mullis in 1985), in order words amplifical of a
target sequenc by mean of the primers (short oligo nucleotides) which bind to the
complementary sequence on the DNA and produce cycle number a large number
of topics of the largest sequence
Steps - Denaturalion (94
o
c fr Imin) dsna -ssona
nnealing (55-6o
o
c 1 min) 1 min/kb final limited
Erotension (72
0
c fr 1 min) prime amended product camplion
19. What is site directed mutagenesis? What is its importance? Explain
Site directed mutagenesis:- An in-vitro method fs creating a specific mutation
in a known sequence and is typically performed by PCR based methods mainly.
Oligonucleotide Traditional PCR (An involvement of a mutated
directed mutagenesis Primer extension primer having insertion/deletion
Inverse PCR or substitution
Importance:-
It acts as a reliable tool for gene manipulations at a specific site.
Simple method for creating variation not tedious like earler techniques since it
utilize PCR.
It helps in study of gene structure and function as mutant proteins are fsmed.
In protein engineering to enhance enzyme specifically activity, affinity to
substrate, stability and mode of action improvement.
Enabled the codon biasing problem solvation which yielded better resulting
promoters in genetic engineering for transgenics formation
20 Describe EMP pathway? How it is regulate?



10 Markes - 6 Qns
1. What are transgenics? How there can help in crop improvement (a0 marks)


2. What are the various vectors used in Cloning? Describe


3. What is T-DNA tagging. Describe with example.


4. What is chloroplast transformation? What is its advantage & limitation
Chloroplast transformation refers to the transfer of gene of interest or entire gene
cassette through site specific recombination from chloroplast vector system to the
chloroplast gsame, resulting in Transformed chloroplast either by direct gene
dividery/biolistic gene approache/PEG mediated method.
Site specific recombination at IRASIRB. site with trnI and trnA of chlonoplast
vector constrceat (using Biolistic gene approach transfer occurs)
Advantages
No position effect (Hetero chranetization leading to gene silencing)
Trans gene containment (Pollen not a problem)
Higher expression (TSS) - greater than 40% protein produced
Site specific and single integration leading to multiple chloropl A transformalian
with entire Biochemical pathway inroduced eq. in carotenoids (Gene-stackery).
Post translational modification possible.
Better expression bacterial genes than nuclear
Disadvantages
High death rate and low regeneration frequency with Biolistic gud approach.
Multiple copy inserted at the same site, deletions it incorporated in DR side and
inverted side if then inversions occurs
Low expression of transgene if Eukaryster in prokeryothi system
Low transformation efficiency
regenration protocol stand required.
5. What is a genome? Describe Dangeros Dequencing what advantage
capillang based dangin sequecing has over traditions one.
Genome:- A complete set of chromosames that one prosent in a gametophytic
cell/ Haproid is called the genome, in order words denoted b n, For maize 2n=2o
and n=10. Is 10 linkage groups (10 chromosames) - comprise whole genome
Sangers sequencing:- Dideoxy termination sequencing method given by sanger
in 1977 used to sequence for the first time 5386 bp (0x174) phage. (M13 phage as
sequencing vector).
Dideoxy nucleotids having lack of 3'0 h group are nable to extend the growing
chain of the nuclotides, when a single strand with 5 ended primer label is extended
by addition of 3d NTP's + 1dd NTP. in each tube termination of nucleotide occurs
and many terminated sequences are obtained as laid in 4 lane get.
Hence that primer used is radiolabelled p32) and through autoradiophgh the
sequence is riad from the base of get to the upmost region. (Read sequences in
sanga method 500-800 bp)
Automation in sanger sequencing method lead to transfer gel based/slab
By fluoresence labelling
4 lane to single lane (Rather than primer labeled)
Capillary Electrophons has (50 mm) size of (Silicon capillary)
Automation single ddNTP moves is detected by CCD camera/photodiode
More precise detection and more no of sequences sequence daily.
Utilization of cycle sequencing method where through PCR based synthe of
template for sequencing da.
6. Which vector you will use for cloning fragments of human DNA & why?
What the advantage your vector of choice have over other one? Explain


SET - 2
1. Bacillus is the example of Naturaly competent micro organism.

2. Define Co incidence & Interference?
Coincidence is the frequency of occurence of a crossing over at positions 1 and 2.
While interference is the ability of an crossing over occuring at position 1 to
influence cross over at nearby postion 2.
either (+ve) or (-ve) influence.
Hence categorized as (+ve) interference or (-ve) interference
3. RFLP was the Ist marker developed in 1980 by Bostein et. al.

4. CAPS is Co-dominout (dominant/codominant) & also called PCR-RFLP.

5. 2% Crossing over = 1% recombination

6. SSRs are found due to Replication Slippage & Unequal crossing over.

7. TFIIH factor onwinds DNA at promoter during transcription in Eukaryotes.

8. TFIIB binds to TBP & recnts Pol II Tf II F Complex in Transcription.
9. DH Nils & RILs are examples of immortal mapping population.
10. Fullform of RAPD is Random aplification of polymophic DNA. & AFLP
Arbitaily fragment length polymupheno.
11. F2/Nils population are used for gene tagging.
12. Size of Tiplasmid is 23Kb & has cloning capacity of 750Kb.
13. pTIC58/Nopoline type Ti plasmid has T-DNA as Dingle transcriptioned unit
& Octopine pTiAcH5 has 2 Independent transcriptioned units.
14. Virc binds to overdrive region in Tiplasmid (Virc, Vird, Vir D2)
15. Vir D1 has Topo isomerase activity & Vir D2 was Endo nuclease activity in
Ti Plasmid transformation process.
16. Southern hybridization was given by E.M. Southern in 1975 & use _____%
Agarose.
17. Western bloting given by ___________ in ____________ yen.
18. Bollguard II has Crylac & Cryzab genes derived from Bt.
19. CRT & DREB one the two examples of Drought tolerant genes.
20. Cycloheximide blocks peptidyl transferase activity nor Eukaryotes.
21. Puronycin acts both in Pro & Eukoryotes & is derieved from mould
Streptomyces alboniger (T/F).
22. Teracycline blocks transcription while steptomycin is a Trisacchande
(disachande/Trisacchand) with binds to 30S submits (50S, 30S, 235)
23. Eukaryotes contains prokaryotic type DNA (true/false)
24. Replication involves RNA primers (true/false)
25. Ist aminoacide addition durint the protein synthesis uses ATP Energy.
26. There are 3 Elongation factors in Protein synthesis of Prokaryotes EF-T4
EF-TS & EF-G
27. Function of If1 is binds to A lite to inhibit binding of aminoacid tRNA at a
site in protein Dyn thesis.
28. Amber = UAG (UAA, UGA, UUA, UAG,)
Ochre = UAA
OPAL = UGA
29. What is polar nutation at one site like pranota results in unexpression of rest
of the genes downstream to the promoter region. Eq. lac gya gens unexpress
in lactor ope if nutation in promote site occurs.
30. C3 plants uses 3 ATP whereas C4 uses 5 ATPs per Glucose molecule.
31. Photorespiration was given by Working & _________.
32. End product of Glycolysis is pyrovate & has 1 3 & 10 as irreversible steps.
33. ___________ type of DNA is longest & __________ type is broadest.
34. Cellulose has __________ bonds where as amylose has ________ bonds.
35. Genome size of E. coli is 4.3 x 10
6
bp & Humans is 3.2 x 10
9
bp.
36. Lines refers to Longr interexpressed nuclear elements sines refers to short
interpressed nuclear element.
37. LK = Wr + TW
L- writs Twia
38. In meta phase stage 10,000 times packed.
39. Eight histones are - H2A H2B H3 H4 with H1 as the linker.
40. Eukayotic cells has type I topoi somerase but type Ii or DNA gyrase is
absent in them (T/F)
5 Marks
1. Elucidate the methods of SNP genotyping.
SNP genotyping methods
- SNP arrays [Hybridization based]
- Dot-blot
- Reverse dot blot
- Micro arrays
Direct sequencing based
overlap -BAC claves
Resequencing of short DNA structure
EST lcone
Enzyme based method
PCR RFLC; primer extension; SI nucleares
Oligo nucldid ligation & mapping of restriction invarus cleavage
2. What are the approaches of/genes for qualitative trauts? Briefly explain



3. Define MAS? What is its advantage & limitation?
Marker assisted selection:- It is a process of determining the location/heridite
of a trial from generate to generate it. It is process whereby a marker &
morpholyical. Brochemical; or DNA based) is used for indirection selecting the
genetic determinant of a traint of interest (disease raistane; productiveity trust
quality etc.
Advantages:- Early detection of GOI Hrangene intro grasim at seeding stage.
Heterzygotes identified
For recognize traits, identify the plant with derived alule and hence avoids selfing
for detective (time saving)/
Coa-effective and labour saving
No Biosafety issues
Not influenced by environment i.e. DNA based markers
No used to create artificial environment fs deletion
Disadvantage- Justial- investment is high.
4. What are the steps for construction of high density linkage map?
Steps involved are:- Selection of cantrasting parents 11 x 111
Divergent/Geographically distinct
Prepare cross
Generation of Mappign population
RILS, NILs, DH, F2; F2;3, BC
Size of population
Genotyping of the mapping population
5000 individual for fine mapping either from heterologoes
Identification of informative markers
Segregation studies
Avoid spurious linkage co-segregation b/w mankut tract the deed
(Statistical leos)
KOSambhi function
LOD score > 3
Assigning linkage groups to the chromosaves.
Construction of high density linkage map.
Note increasing the population size increase rare recombinant chances (2%
crossing over = 1% recabinet).
5) What are the steps of map based cloning? What are its advantage &
limitation.
Map based cloning
Identify that clone with both markers (M1 & M2)
that flank the GOI
prepare primers from marker and PCR. each PCR gives a clone
Select the clone with both the markers
Advantages
Direct PCT based method to land into In silvio approach
BIAST obtain possible candidate genes
Disadvantages
Not efficient for Long exams bearing genes with intronic sequences
What advantages caps marker have over RFLP
RFLP and CAPS both involve R.E digesters and Hybridization with probe to
identify the revied sequence