CHAPTER 6
CHAPTER 6
1 Define transformation in Griffith's experiment. Discuss how it helps in the identification of DNA as the
genetic material.
2 Who revealed biochemical nature of the transforming principle? How was it done?
3 Discuss the significance of heavy isotope of nitrogen in the Meselson and Stahl's experiment.
4 Define a cistron. Giving examples differentiate between monocistronic and polyeistronic transcription unit.
5 Give any six features of the human genome.
6 During DNA replication, why is it that the entire molecule does not open in one go? Explain replication fork.
What are the two functions that the monomers (d NTPs) play?
7 Retroviruses do not follow central Dogma. Comment.
8 In an experiment, DNA is treated with a compound which tends to place itself amongst the stacks of
nitrogenous base pairs. As a result of this, the distance between two consecutive base increases. from 0.34
nm to 0.44 nm calculate the length of DNA double helix (which has 2 ×109 bp ) in the presence of saturating
amount of this compound.
9 What would happen if histones were to be mutated and made rich in acidic amino acids such as aspartic
acid and glutamic acid in place of basic amino acids such as lysine and arginine?
10 Recall the experiments done by Frederick Griffith, Avery, MacLeod and McCarty, where DNA was
speculated to be the genetic material. If RNA, instead of DNA was the genetic material, would the heat killed
strain of Pneumococcus have transformed the R-strain into virulent strain? Explain.
11 You are repeating the Hershey-Chase experiment and are provided with two isotopes: 32 P and 15 N (in
place of 35 S in the original experiment). How do you expect your results to be different?
12 There is only one possible sequence of amino acids when deduced from a given nucleotides. But multiple
nucleotides sequence can be deduced from a single amino acid sequence. Explain this phenomena.
13 A single base mutation in a gene may not 'always' result in loss or gain of function. Do you think the
statement is correct? Defend your answer.
14 A low level of expression of lac operon occurs at all the time. Can you explain the logic behind this
phenomena.
15 How has the sequencing of human genome opened new windows for treatment of various genetic
disorders. Discuss amongst your classmates.
16 The total number of genes in humans is far less (¿ 25,000) than the previous estimate (upto 1,40,000 gene).
Comment.
17 Now, sequencing of total genomes getting is getting less expensive day by the day. Soon it may be
affordable for a common man to get his genome sequenced. What in your opinion could be the advantage
and disadvantage of this development?
18 Would it be appropriate to use DNA probes such as VNTR in DNA finger printing of a bacteriaphage?
19 During in vitro synthesis of DNA, a researcher used 2, 3' - dideoxy cytidine triphosphate as raw nucleotide
in place of 2'-deoxy cytidine. What would be the consequence?
20 What background information did Watson and Crick have made available for developing a model of DNA?
What was their contribution?
21 What are the functions of (i) methylated guanasine cap, (ii) poly-A "tail" in a mature on RNA?
22 Do you think that the alternate splicing of exons may enable a structural gene to code for several
isoproteins from one and the same gene? If yes, how? If not, why so?
23 Comment on the utility of variability in number of tandem repeats during DNA finger printing.
1 Give an account of Hershey and Chase experiment. What did it conclusively prove? If both DNA and
proteins contained phosphorus and sulphur do you think the result would have been the same?
2 During the course of evolution why DNA was chosen over RNA as genetic material? Give reasons by first
discussing the desired criteria in a molecule that can act as genetic material and in the light of biochemical
differences between DNA and RNA.
3 Give an account of post transcriptional modifications of a eukaryotic mRNA.
4 Discuss the process of translation detail.
5 Define an operon. giving an example, explain an Inducible operon.
6 'There is a paternity dispute for a child'. Which technique can solve the problem. Discuss the principle
involved.
7 Give an account of the methods used in sequencing the human genome.
8 List the various markers that are used in DNA finger printing.
9 Replication was allowed to take place in the presence of radioactive deoxynucleotides precursors in E.coli
that was a mutant for DNA ligase. Newly synthesised radioactive DNA was purified and strands were
separated by denaturation. These were centrifuged using density gradient centrifugation. Which of the
following would be a correct result?
Answers and Solutions
MULTIPLE-CHOICE QUESTIONS :
1. b
2. c
3. c
4. c
5. c
6. b
7. d
8. d
9. d
10. b
11. c
12. d
13. b
14. b
15. b
16. b
17. d
18. a
19. d
20. b
21. c
22. d
23. a
24. c
25. b
26. b
27. a
28. a
1. In the experiment both the scientists used the bacteriophage which are composed of DNA and proteins,
infected bacteria are inserted into the host bacterial cell walls but after the experiment not get desired
results. DNA is a hereditary material after the subsequent discoveries and interpretation all serve to prove.
And in 1969 Hershey shared the Nobel Prize in Physiology and Medicine with the Scientist Max.
In the experiment Hershey and Chase examined the different phages and isolated the phages in the
subsection. The labelling of each different element is called an isotope obtained from the viruses that are
composed from the protein shell and DNA and both are analyzed differently. Phosphorus not containing
amino acid but containing DNA, T-2 phage is labelled by using the radioactive phosphorus-32. After that
kept the media growing for 4 hours. And obtained the progeny from the infected bacteriophage bacteria
that contained radioactive isotopes structure. In this sulfur labelled phages and once for phosphorus
labelled phages are used. And after the final concluded results the DNase is protected by the DNA. And
proved DNA is the source of genetic material not the RNA.
2. A molecule that can act as a genetic material must fulfil the following
(i) It should be able to generate its replica (replication).
(ii) It should chemically and structurally be stable.
(iii) It should provide the scope for slow changes (mutation) that are required for evolution.
(iv) It should be able to express itself in the form of Mendelian.
Biochemical differences between DNA and RNA
(i) Both nucleic acid (DNA and RNA) are able to direct their duplication proteins fails for the
first criteria.
(ii) RNA is reactive, it also acts are catalyst, hence DNA is less reactive and structurally
more stable than RNA.
(iii) Presence of thymine at the place of uracil also confers additional stability to DNA.
3. The primary transcripts (hn-RNA) contain both the exons and the introns and are non-functional. Hence, it
is subjected to a process called splicing where the introns are removed and exons are joined in a defined
order. Intron is the portion of gene which is transcribed but not translated. In prokaryotes hnRNA is absent
so splicing in not required. hnRNA undergoes additional processing called as capping and tailing. In
capping an unusual nucleotide (methyl guanosine triphosphate) is added to the 5′-end of hnRNA. In tailing,
adenylate residues (200-300) are added at 3′-end in a template independent manner. It is the fully
processed hnRNA, . now called mRNA, that is transported out of the nucleus for translation.
Amino acids are added one by one, translated into polypeptide sequences dictated by DNA and represented
by mRNA.
At the end, a release factor binds to the stop codon, terminating translation and releasing the complete
polypeptide from the ribosome
5. The concept of operon was first proposed in 1961, by Jacob and Monad. An operon is a unit of prokaryotic
gene expression which indludes coordinately gene product.
Components of an Operon
(i) Structurel of an gene The fragment of DNA which transcribe mRNA for polypeptide synthesis.
(ii) Promoter The sequence of DNA where RNA polymerase binds and initiates transcriptin of structural
genes is called promoter.
(iii) Operator The sequence of DNA adjacent to promoter where specific repressor protein binds is called
operator.
(iv) Regulator gene The gene that codes for the repessor protein that binds to the operator and suppresses
its acitvity as a result of which transcription will be switched off .
(v) Inducer The substrate that prevents the repressor from binding to the operator, is called an inducer. As
a result transcription is switched on. It is a chemical of diverse nature like metabolite, hormone substrante,
etc.
Inducible Operon System
An inducible operon system is a regulated unit of ganetic material which is switched on in response ot the
presence of a chemical.e.g., the lactose or lac-operon of E.coli.
The lactose operon The lac z,y,a genes are transcribed from a lac transcription unit uner the control of a
single promoter. They encode enzyme required for the use of lactose as a carbon source. The lac i gene
product, the lac repressor, is expressed from a separate transcription unit upstream from the operator .
lac operon consists of three structural genes (z,y and a ),operator and a separate regulatory gene.
The three structural genes (a,y and a) transcribe a polycistronic mRNA.
Gene z codes for bets-galactossidase(β−gal) enzyme which breaks lactose into galactose and glucose.
Gene y codes for permease, which incrreases the permeability of the cell to lactose .
Gene a codes for enzyme transacetyiase, which catalyses the transacetylation of lactose in
When Lactose is Absent
(i) When lactose is absent, i gene regulates and produces repressor mRNA which translate repression .
(ii) The operon is switched off.
when Lactose is Present
(i) Lactose acts as an inducer which binds to the repressor and an inactive repressor.
(ii)The repressor fails to bind to the oparator region .
(iii)The RNA polymerase binds to the operator and transcript lac mRNA.
(iv) lac mRNA is polycistronic,i.e., produces all three enzymes , β-galactosidase, permease and
transscatylase.
(v) The lac operon is switched on .
6. DNA finger printing is used to solve the paternity dispute. DNA fingerprinting involves identifying
differences in some specific regions sequence called as repetitive DNA, because in these sequences, a small
stretch of DNA is repeated many times. These repetitive DNA are separated from bulk genomic DNA as
different peaks during density gradient centrifugation.
The bulk DNA forms a major peak and the other small peaks are referred to as satellite DNA. Depending on
base composition (A : T rich or G :C rich), length of segment, and number of repetitive units, the satellite
DNA is classified into many categories, such as micro- satellites, mini-satellites etc. These sequences
normally do not code for any proteins, but they form a large portion of human genome.
These sequence show high degree of polymorphism and form the basis of DNA fingerprinting. Since DNA
from every tissue (such as blood, hair- follicle, skin, bone, saliva, sperm etc.), from an individual show the
same degree of polymorphism, they become very useful identification tool in forensic applications. Further,
as the polymorphisms are inheritable from parents to children, DNA fingerprinting is the basis of paternity
testing, in case of disputes.
The technique of DNA fingerprinting was initially developed by Alec Jeffreys. Lalji Singh is called father of
Indian DNA fingerprinting or DNA profiling or DNA typing. He used a satellite DNA as probe that shows
very high degree of polymorphism. It was called as Variable Number of Tandem Repeats (VNTR).
The technique, as used earlier, involved Southern blot hybridisation using radiolabelled VNTR as a probe. It
included
(i) Isolation of DNA, (ii) Digestion of DNA by 8 restriction endonucleases, (iii) Separation of DNA fragments
by electrophoresis, (iv) Transferring (blotting) of separated DNA fragments to synthetic membranes, such
as nitrocellulose or nylon, (v) Hybridisation using labelled VNTR probe, and (vi) Detectionof hybridised
DNA fragments by autoradiography.
7. Sequencing of human genome has made it possible to understand the link between various genes and their
functions. If there are any gene defects that express as disorders or that increase the susceptibility of an
individual to a disease then specific gene therapies can be worked out
Methodologies of human genome sequencing
The methods involve two major approaches
(i) Expressed Sequence Tags (ESTs) This method focusses on identifying all the genes that are expressed
as RNA.
(ii) Sequence annotation It is an approach of simply sequencing the whole set of genome that contains all
the coding and non-coding sequences, and later assigning different regions in the sequence with functions.
For sequencing, first the total DNA from cell is i.e., solated and broken down in relatively small sizes as
fragments.
There DNA fragments are cloned in suitable host using suitable vectors. When bacteria is used as vector,
they are called Bacterial Artificial Chromosomes (BAC) and when yeast is used as vector, they are called
Yeast Artificial Chromosomes (YACs).
Frederick Sanger developed a principle according to which the fragments of DNA are sequenced by
automated DNA sequences.
On the basis of overlapping regions on DNA fragments, these sequences are arranged accordingly. For
alignment of these sequences, specialised computer-based programmes were developed. Finally, the
genetic and physical maps of the genome were constructed by collecting information about certain
repetitive DNA sequences and DNA polymorphism, based on endonuclease recognition sites.
8. (i) Restriction endonucleases.
(ii) Fluorescent dyes for separating VNTRs or RFLPs.
(iii) X-ray films.
(iv) Satellite DNAs.
(v) Southern blotting.
(vi) Hybridised VNTRs.
(vii) Radioactive DNA probes.
9. In above case, as E.coli is a mutant for DNA ligase, it will result in no further joining of Okazaki fragments on
lagging strand.
This will ultimately result into the formation of both high molecular weight fragments (on leading strands)
and low molecular weight fragments (on lagging strand). Hence, only the graph (a) could be the
appropriate result after centrifugation.