CHAPTER 6

MOLECULAR BASIS OF INHERITANCE

MULTIPLE-CHOICE QUESTIONS

1 In a DNA strand the nucleotides are linked together by:

a. glycosidic bonds
b. phosphodiester bonds
c. peptide bonds
d. hydrogen bonds
2 A nucleoside differs from a nucleotide. It lacks the:
a. base
b. sugar
c. phosphate group
d. hydroxyl group
3 Both deoxyribose and ribose belong to a class of sugars called:
a. trioses
b. hexoses
c. pentoses
d. polysaccharides
4 The fact that a purine base always pairs through hydrogen bonds with a pyrimidine base in the DNA double
helix leads to:
a. the antiparallel nature
b. the semiconservative nature
c. uniform width throughout DNA
d. uniform length in all DNA
5 The net electric charge on DNA and histones is:
a. both positive
b. both negative
c. negative and positive, respectively
d. zero
6 The promoter site and the terminator site for transcription are located at:
a. 3' (downstream) end and 5' (upstream) end, respectively of the transcription unit
b. 5' (upstream) end and 3' (downstream) end, respectively of the transcription unit
c. the 5 ' (upstream) end
d. the 3 ' (downstream) end
7 Which of the following statements is the most appropriate for sickle cell anaemia?
a. It cannot be treated with iron supplements
b. It is a molecular disease
c. It confers resistance to acquiring malaria
d. All of the above
8 Which of the following is true with respect to AUG?
a. It codes for methionine only
b. It is an initiation codon
c. It codes for methionine in both prokaryotes and eukaryotes
d. All of the above
9 The first genetic material could be:
a. protein
b. carbohydrates
 c. DNA
d. RNA
10 With regard to mature mRNA in eukaryotes:
a. exons and introns do not appear in the mature RNA
b. exons appear but introns do not appear in the mature RNA
c. introns appear but exons do not appear in the mature RNA
d. both exons and introns appear in the mature RNA
11 The human chromosome with the highest and least number of genes in them are respectively:
a. Chromosome 21 and Y
b. Chromosome 1 and X
c. Chromosome 1 and Y
d. Chromosome X and Y
12 Who amongst the following scientists had no contribution in the development of the double helix model for
the structure of DNA?
a. Rosalind Franklin
b. Maurice Wilkins
c. Erwin Chargaff
d. Meselson and Stahl
13 DNA is a polymer of nucleotides which are linked to each other by 3-5' phosphodiester bond. To prevent
polymerisation of nucleotides, which of the following modifications would you choose?
a. Replace purine with pyrimidines
b. Remove/Replace 3' OH group in deoxy ribose
c. Remove/Replace 2' OH group with some other group in deoxy ribose
d. Both 'b' and 'c'
14 Discontinuous synthesis of DNA occurs in one strand, because:
a. DNA molecule being synthesised is very long
b. DNA dependent DNA polymerase catalyses polymerisation only in one direction ( 5' →3' )
c. it is a more efficient process
d. DNA ligase joins the short stretches of DNA
15 Which of the following steps in transcription is catalysed by RNA polymerase?
a. Initiation
b. Elongation
c. Termination
d. All of the above
16 Control of gene expression in prokaryotes take place at the level of:
a. DNA-replication
b. Transcription
c. Translation
d. None of the above
17 Which of the following statements is correct about the role of regulatory proteins in transcription in
prokaryotes?
a. They only increase expression
b. They only decrease expression
c. They interact with RNA polymerase but do not affect the expression
d. They can act both as activators and as repressors
18 Which was the last human chromosome to be completely sequenced:
a. Chromosome 1
b. Chromosome 11
c. Chromosome 21
d. Chromosome X
19 Which of the following are the functions of RNA?
a. It is a carrier of genetic information from DNA to ribosomes synthesising polypeptides.
b. It carries amino acids to ribosomes.
c. It is a constituent component of ribosomes.
d. All of the above.
20 While analysing the DNA of an organism a total number of 5386 nucleotides were found out of which the
proportion of different bases were: Adenine ¿ 29 % , Guanine ¿ 17 % , Cytosine ¿ 32 %, Thymine ¿ 17 % .
Considering the Chargaff's rule it can be concluded that:
a. it is a double stranded circular DNA
b. It is single stranded DNA
c. It is a double stranded linear DNA
d. No conclusion can be drawn
21 In some viruses, DNA is synthesised by using RNA as template. Such a DNA is called:
a. A-DNA
b. B-DNA
c. cDNA
d. rDNA
22 If Meselson and Stahl's experiment is continued for four generations in bacteria, the ratio of
15 15 15 14 14 14
N / N : N / N : N / N containing DNA in the fourth generation would be:
a. 1 :1:0
b. 1 :4 : 0
c. 0 :1 :3
d. 0 :1 :7
23 If the sequence of nitrogen bases of the coding strand of DNA in a transcription unit is:
' '
5 − AT G AAT G - 3 ,
the sequence of bases in its RNA transcript would be;
a. 5' - A U G A A U G - 3'
b. 5' - U A C U U A C - 3'
c. 5' - C A U U C A U - 3'
d. 5' - G U A A G U A - 3'
24 The RNA polymerase holoenzyme transcribes:
a. the promoter, structural gene and the terminator region
b. the promoter and the terminator region
c. the structural gene and the terminator region
d. the structural gene only.
25 If the base sequence of a codon in mRNA is 5' -AUG-3', the sequence of tRNA pairing with it must be:
a. 5' - UAC - 3'
b. 5' - CAU - 3'
c. 5' - AUG - 3'
d. 5' - GUA - 3'
26 The amino acid attaches to the tRNA at its:
a. 5' - end
b. 3' - end
c. Anti codon site
d. DHU loop
27 To initiate translation, the mRNA first binds to:
a. The smaller ribosomal sub-unit,
b. The larger ribosomal sub-unit
c. The whole ribosome
d. No such specificity exists.
28 In E.coli, the lac operon gets switched on when:
a. lactose is present and it binds to the repressor
b. repressor binds to operator
c. RNA polymerase binds to the operator
d. lactose is present and it binds to RNA polymerase

VERY SHORT ANSWER TYPE QUESTIONS

1 What is the function of histones in DNA packaging?

2 Distinguish between heterochromatin and euchromatin. Which of the two is transcriptionally active?
3 The enzyme DNA polymerase in E.coli is a DNA dependent polymerase and also has the ability to proof-
read the DNA strand being synthesised. Explain. Discuss the dual polymerase.
4 What is the cause of discontinuous synthesis of DNA on one of the parental strands of DNA? What happens
to these short stretches of synthesised DNA?
5 Given below is the sequence of coding strand of DNA in a transcription unit
3 'A ATG C A G C TATTAGG-5'
write the sequence of
a) its complementary strand
b) the mRNA
6 What is DNA polymorphism? Why is it important to study it?
7 Based on your understanding of genetic code, explain the formation of any abnormal hemoglobin molecule.
What are the known consequences of such a change?
8 Sometimes cattle or even human beings give birth to their young ones that are having extremely different
sets of organs like limbs/position of eye(s) etc. Comment.
9 In a nucleus, the number of ribonucleoside triphosphates is 10 times the number of deoxy x10
ribonucleoside triphosphates, but only deoxy ribonucleotides are added during the DNA replication.
Suggest a mechanism.
10 Name a few enzymes involved in DNA replication other than DNA polymerase and ligase. Name the key
functions for each of them.
11 Name any three viruses which have RNA as the genetic material.

SHORT ANSWER TYPE QUESTIONS

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.

LONG ANSWER TYPE QUESTIONS

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

VERY SHORT ANSWER TYPE QUESTIONS

1. Functions of histones in DNA packaging are

(i) Histones as units of octamer participate in primary packaging of DNA.
(ii) Basic histone proteins neutralise the acidic DNA molecule.
2. 1. Euchromation : Regions of chromatin that are loosely packed and stained lightly. It is transcriptionally
active.
2. Heterochromation : Regions of chromatin that are densely packed and stained dark. It is transcriptionally
inactive.
3. In bacteria, three types of DNA polymerases are there. All of them can add nucleotides in 5’  3’ direction.
They process exonuclease activity as well. DNA polymerase III can proofread the newly synthesised strand
and senses the wrong base insertions. It deletes wrong bases and helps correct the mistake by putting in
the right one, DNA polymerase. The only mistake it cannot corrects substitution of uracil in place of thymin.
It can repair any damages done to DNA by UV exposure, etc., or the left over proofreading mistakes. It
detects mutation caused by UV, removes mismatched pairs and puts back the right ones.
4. There is discontinuous synthesis of DNA on the lagging strand because the DNA polymerase moves from 5’
to 3’ direction only.
The lagging strand runs from 3’ to 5’ direction.
So, by looping back mechanisms small stretches of DNA are synthesized.
The short stretches of synthesized DNA called okazaki fragments are later joined together by DNA ligase.
5. According to base complementary rules,
(a) 5'TTACGTCGATAATCC-3'
(b) 5'CGAUUAUCGACGUAA-3'
RNA uses the base uracil (U) rather than thymine (T). So, in RNA the base pairs are
Adenine (A) pairs with uracil (U)
Guanine (G) pairs with cytosine (C).
6. DNA polymorphism refers ro the variation in DNA arising through mutation at non-coding sequences.
A special type of polymorphism, called VNTR (Variable Number of Tendem Repeats), is composed of
repeated copies of a DNA sequence that lie adjacent to one another on the chromosome. Since,
polymorphism is the basis of genetic mapping of humen genome, therefore, it forms the basis of DNA
fingerprinting too. history as well as in cese of paternity testing .
7. Due to point mutation in b-globin chain of haemoglobin molecule, glutamic acid (Glu) is replaced by valine
(Val) at the sixth position. Under stress condition erythrocytes lose their circular shape and become sickle-
shaped. As a result, the cells cannot pass through narrow capillaries. Blood capillaries are clogged and
thus affect blood supply to different organs.
8. There is a disturbance in co-ordinated regulation of expression of sets of genes associated with organ
development.
9. DNA polymerase is highly specific to recognise only deoxy ribonucleoside triphosphates. Therefore it
cannot hold RNA nucleotides.
10. (i) helicase – opens the helix
(ii) topoisomerases – removes the supercoiling of DNA
(iii) Primase: synthesises RNA primer
(iv) Telomerase: to synthesises the DNA of telomeric end of chromosomes.
11. In some viruses, RNA is the genetic material. e.g., Tobacco mosaic viruscs, QB bacteriophage, HIV, influenza
virus, etc.

SHORT ANSWER TYPE QUESTIONS

1. In Griffith’s experiment, transformation can be defined as a change in the genetic constitution of an

organism by picking out up DNA from the environment (from dead organisms).
Transformation helps in identification of DNA as a genetic material. When heat was used to kill the virulent
bacteria, they died but not their genetic material (DNA). This DNA when picked up by non-virulent bacteria
made them capable of causing infection. Since, ability to cause infection could be passed on by these
organisms to their progeny, it was concluded that DNA was the material that was inherited.
2. Oswald, Avery, Colin MacLeod and Maclyn McCarty revealed biochemical nature of the transforming
principle. They reported Griffith’s experiment in an in vitro system in order to determine biochemical
nature of transforming principle.
They reported that DNA from the heat-killed S-type bacteria caused the transformation of non-virulent R-
type bacteria into virulent S-type bacteria. They also discovered that proteases and RNase did not affect
transformation while DNase inhibited the process. They concluded that DNA is the hereditary material.
3. They performed experiments on E. coli to prove that DNA replication is semi-conservative. They first grew
the bacteria in a medium containing 15NH4Cl (in which 15N is the heavy istope of nitrogen) for many
generations.
 Then they transferred the cells into a medium with normal 14NH4 Cl (in which 14N is the lighter isotope) and
took the samples at various definite time intervals as the cells multiplied. The extracted DNAs were
centrifuged and measured to get their densities. The DNA extracted from the culture after one generation of
transfer from then 15N mediun to 14N mediun, (i.e., after 20 minutes E.coli divides every 20 minutes)
showed an intermediate hybrid density, i.e., both heavy and light nitrogen, which proved the semi-
conservative nature of DNA.
4. A cistron is stretch of base sequences that codes for one polypeptide chain including adjacent control
regions. It may also code for a tRNA, rRNA molecule or may perform other specific functions including
regulating functions of other cistrons. This term has replaced the definition of a gene. Monocistronic
transcription unit will have all the regulatory and coding sequences for a single polypeptide, whereas
polycistronic may have coding sequences for more than one polypeptide. In eukaryotic cells almost all the
messenger RNAs are monocistronic. In prokaryotes, lac operon coding sequence would be an example of
polycistronic DNA region.
5. Salient features of human genome
(i) The human genome contains 3164.7 million nucleotide bases.
(ii) The average gene consists of 30000 the largest know human gene being dystrophin at
2.4 Million bases.
(iii) The total number of genes is estimated to be 30000 and 99.9% nucleotide bases are
exactly the same in all people.
(iv) The functions are unknown for over 50% of the discovered genes.
(v) Less than 2% of the genome codes for proteins.
(vi) The human genome contains large repeated sequences.
6. While replicating, the entire DNA molecule to keep the whole molecule stabilised does not open in one go
because it would be highly expens energetically. Actually unwiding creates tension in the molecule as
uncoiled parts. Actually, unwinding creates tension in the molecule as uncoiled parts start forming super
coils due to the interaction of exposed nucleotides. Instead, helicase enzyme acts on the double strand at ori
site (origin of replication) and a small stretch is unzipped. Immediately, it is held and stabilised by single
strand binding proteins. Slowly with the help of enzymes, exposed strands are copied as a point of
unwinding moves and ahead in both directions.
It gives an appearance of Y-shaped structure which is called replication fork.
The two functions that the monomer units of NTPs play are
(i) They pair up with exposed nucleotides of the template strand and make phosphodiester linkages and
release a pyrophosphate.
(ii) Hydrolysis of this pyrophosphate by enzyme pyrophosphatase releases energy that will facilitate
making hydrogen bonds between free nucleotides and bases of the template strand.
7. Genetic material of retrovirus is RNA. At the time of synthesis of protein, RNA is ‘reverse transcribed’ to its
complementary DNA first, which is opposite to the central dogma. Hence, retrovirus are not known to
follow central dogma.
8. The length of DNA double helix = 2 × 109 × 0.44 × 10− 9 / bp.
9. If histone proteins were rich in acidic amino acids instead of basic amino acids then they may not have any
role in DNA packaging in eukaryotes as DNA is also negatively charged molecule. The packaging of DNA
around the nucleosome would not happen. Consequently, the chromatin fibre would not be formed.
10. RNA is more labile and prone to degradation (owing to the presence of 2’ OH group in its ribose). Hence
heat-killed S-strain may not have retained its ability to transform the R-strain into virulent form if RNA was
its genetic material.
11. Use of 15N will be inappropriate because method of detection of 35p and 15N is different (32p being a
radioactive isotope while 15N is not radioactive but is the heavier isotope of Nitrogen). Even if 15N was
radioactive then its presence would have been detected both inside the cell (15N incorporated as
 nitrogenous base in DNA) as well as in the supernatant because 15N would also get incorporated in amino
group of amino acids in proteins). Hence the use of 15N would not give any conclusive results.
12. Some amino acids are coded by more than one codon (known as degeneracy of codons), hence on deducing
a nucleotide sequence from an amino acid sequence, multiple nucleotide sequence will be obtained. For
e.g., Ile has three codous: AUU, AUC AUA hence a depeptide Met–Ile can have the following nucleotide
sequence:
(i) AUG – AUU
(ii) AUG – AUC
(iii) AUG – AUA
and if, we deduce amino acid sequence the above nucleotide sequences, all the three will code for Met–Ile\
13. The statement is correct. Because of degeneracy of codons, mutations at third base of codon, usually does
not result into any change in phenotype. This is called silent mutations.
14. In the complete absence of expression of lac operon, permease will not be synthesised which is essential for
transport of lactose from medium into the cells. And if lactose cannot be transported into the cell, then it
cannot act as inducers hence, cannot relieve the lac operon from its repressed state.
15. The sequencing of human genome helped in enhancing the basic understanding of genetics and immunity
to various disorders. Various genes that cause genetic disorders were identified with the help of this
project. It was found that more than 1200 genes are responsible for common human cardiovascular
diseases, endocrine diseases (like diabetes), neurological, disorders (like Alzeimer’s disease, cancers and
many more. These diseases can be treated easily by knowing the particular gene responsible for the
particular disease.
16. The total number of genes is estimated at 25000 much lower than previous estimates of 140000 that had
been based on extrapolations from gene-rich areas as opposed to a composite of gene-rich and gene-poor
areas.
Almost all (99.9%) nucleotide bases are exactly the same in all people. Functions for over 50% discovered
genes are not known yet. Scientist have identified about 1.4 million locations where single-base DNA
difference (SNPs or Single Nucleotide Polymorphisms) occur in humans. This information promises to
revolutionise the processes of finding chromosomal locations for disease-associated sequence and tracing
human history.
17. Human genome helps to find out the complete genome sequence of the human. It has many advantages and
disadvantages.
Some important advantages
It provides the knowledge of the effects of variations of DNA among individuals can revolutionise the ways
to diagnose, treat and prevent many diseases that affect humans. It also provides clues to the
understanding of human biology. It helps to find out the human evolution. Identification through DNA
forensics is also possible.
Some important disadvantages
People might discover and untreatable genetic disease. People may abuse the knowledge obtained from the
HGP. Problem can occur for the ownership of the genetic test result and the patenting of human genes and
DNA. People believe that they are special and unique in their own ways and may wish to remain like that.
18. Bacteriaphage does not have repetitive sequences such as VNTRs in its genome as its genome is very small
and have all the coding sequence. DNA finger printing is not done for phages.
19. Further polymerisation would not occur, as the 3¢ OH on sugar is not there to add a new nucleotide for
forming ester bond.
20. Wastson and Crick had the following informations which helped them to develop a model of DNA.
(i) Chargaffs’ Law suggesting A = T, and C = G.
(ii) Wilkins and Rosalind Franklin’s work on DNA crystal’s X-ray diffraction studies about DNA’s physical
structure.
 (iii) Watson and crick proposed
a. How complementary bases may pair
b. Semi conservative replication and
c. Mutation through tautomerism.
21. The function of Methylated Guanosine Cap: It regulates nuclear export of mRNA. It promotes translation.
(Fully processed hnRNA Is called mRNA).
The function of Poly-A Tail: Protects RNA from degradation by exonucleases. Plays an important role in
transcription termination.
22. Functional mRNA of structural genes need not always include all of its exons. This alternate splicing of
exons is sex-specific, tissue-specific, and even developmental stage-specific. By such alternate splicing of
exons, a single gene may encode for several isoproteins and/or proteins of similar class. In absence of such
a kind of splicing, there should have been new genes for every protein/isoprotein. Such an extravagancy
has been avoided in natural phenomena by way of altemate splicing.
23. Tandemness in repeats provides many copies of the sequence for finger-printing and variability in nitrogen
base sequences present in them. Being individual-specific, this proves to be useful in the process of DNA
fingerprinting.

LONG ANSWER TYPE QUESTIONS

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.

4. Translation refers to the process of polymerisation of amino acids to form a polypeptide.

The order and sequence of amino acids are defined by the sequence of bases in the mRNA.
The amino acids are joined by a bond which is known as a peptide bond.
Formation of a peptide bond requires energy.
Therefore, in the first phase itself amino acids are activated in the presence of ATP and linked to their
cognate tRNA - a process commonly called as charging of tRNA or aminoacylation of tRNA to be more
specific.
If two such charged tRNAs are brought close enough the formation of peptide bond between them would be
favoured energetically.
The presence of a catalyst would enhance the rate of peptide bond formtion.
The cellular factory responsible for synthesising proteins is the ribosome.
The ribosome consists of structural RNAs and about 80 different proteins.
In its inactive state, it exists as two subunits, a large subunit and a small subunit.
When the small subunit encounters an mRNA, the process of translation of the mRNA to protein begins.
There are two sites in the large subunit, for subsequent amino acids to bind to and thus be close enough to
each other for the formation of a peptide bond.
The ribosome also acts as a catalyst (23S rRNA in bacteria is the enzyme ribozyme) for the formation of
peptide bond.
A translational unit in mRNA is the sequence of RNA that is flanked by the start codon (AUG) and the stop
codon and codes for a polypeptide.
An mRNA also has some additional sequences that are not translated and are referred as untranslated
regions. (UTR).
The UTRs are present at both 5.-end (before start codon) and at 3.-end (after stop codon).
They are required for efficient translation process.
For initiation, the ribosome binds to the mRNA at the start codon (AUG) that is recognised only by the
initiator tRNA.
The ribosome proceeds to the elongation phase of protein synthesis.
During this stage, complexes composed of an amino acids linked to tRNA, sequentially bind to the
 appropriate codon in mRNA by forming complementary base pairs with the tRNA anticodon.
The ribosome moves from codon to codon along the mRNA.

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.