Molecular Basis of Inheritance NCERT HIGHLIGHT by SEEP Pahuja
Molecular Basis of Inheritance NCERT HIGHLIGHT by SEEP Pahuja
Molecular Basis of Inheritance NCERT HIGHLIGHT by SEEP Pahuja
dingns*ERI
you
CHAPTER 5
MOLECULAR BASIS OF
INHERITANCE
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delinear Bacteriophage lambda has 48502 base pairs (bp), Escherichia coli has
4.6 × 106 bp, and haploid content of human DNA is 3.3 × 109 bp. Let us
de circula
genome Dippid=6.8x109bp.
->
tuposition.
U- I
5
Let us recapitulate the chemical structure of a polynucleotide chain (DNA
or RNA). A nucleotide has three components – a nitrogenous base, a
pentose sugar (ribose in case of RNA, and deoxyribose for DNA), and a
5-Methyl
Uracil
phosphate group. There are two types of nitrogenous bases – Purines
(Adenine and Guanine), and Pyrimidines (Cytosine, Uracil and Thymine).
Cytosine is common for both DNA and RNA and Thymine is present in
DNA. Uracil is present in RNA at the place of Thymine. A nitrogenous
base is linked to the OH of 1' C pentose sugar through a N-glycosidic
linkage to form a nucleoside, such as adenosine or deoxyadenosine,
guanosine or deoxyguanosine, cytidine or deoxycytidine and uridine or
deoxythymidine. When a phosphate group is linked to OH of 5' C of a
nucleoside through phosphoester linkage, a corresponding nucleotide
(or deoxynucleotide depending upon the type of sugar present) is formed.
Two nucleotides are linked through 3'-5' phosphodiester linkage to form
a dinucleotide. More nucleotides can be joined in such a manner to form
a polynucleotide chain. A polymer thus formed has at one end a free
80
(n=nucleotide)
Figure 5.1 A Polynucleotide chain
my phosphodiestelbond:
n-1
186 ·lineal
dS. DNA. =
2n-2
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X-Ray
crystallography
proposed a very simple but famous Double Helix model for the structure
of DNA. One of the hallmarks of their proposition was base pairing between
t
the two strands of polynucleotide chains. However, this proposition was
also based on the observation of Erwin Chargaff that for a double stranded
Sawtelical sty
andsDNAG
-
DNA Pu +
By.
DNA, the ratios between Adenine and Thymine and Guanine and Cytosine
are constant and equals one. ↳ only
=
the sequence of bases in one strand is known then the sequence in other cG =
strand can be predicted. Also, if each strand from a DNA (let us call it as a
parental DNA) acts as a template for synthesis of a new strand, the two
double stranded DNA (let us call them as daughter DNA) thus, produced
would be identical to the parental DNA molecule. Because of this, the genetic
implications of the structure of DNA became very clear.
The salient features of the Double-helix structure of DNA are as follows:
(i) It is made of two polynucleotide chains, where the backbone is
constituted by sugar-phosphate, and the bases project inside.
(ii) The two chains have anti-parallel polarity. It means, if one
chain has the polarity 5' 3' , the other has 3 ' 5 ' .
(iii) The bases in two strands are paired through hydrogen bond
(H-bonds) forming base pairs (bp). Adenine forms two hydrogen
bonds with Thymine from opposite strand and vice-versa.
Similarly, Guanine is bonded with Cytosine with three H-bonds.
As a result, always a purine comes opposite to a pyrimidine. This 81
generates approximately uniform distance between the two
strands of the helix (Figure 5.2).
(iv) The two chains are coiled in a right-handed fashion. The pitch
of the helix is 3.4 nm (a nanometre is one billionth of a
metre, that is 10-9 m) and there are roughly 10 bp in each
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8
Hallmark Watson crick
↑
Bestairing
is
complementary
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double
Stability
to
Helical str
①H-Bond
② Bp plane is
same le
y.
proposed the Central dogma in molecular
biology, which states that the genetic
Figure 5.3 DNA double helix information flows from DNARNAProtein.
82
a Central dogma
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not have a defined nucleus, the DNA is not scattered Figure 5.4a Nucleosome
--
throughout the cell. DNA (being negatively charged)
poly-
-
Mbelre
a unit of eight molecules called histone octamer. A1,
The negatively charged DNA is wrapped around the positively charged
histone octamer to form a structure called nucleosome (Figure 5.4 a). A
typical nucleosome contains 200 bp of DNA helix. Nucleosomes constitute
body ach)
> ↓
the repeating unit of a structure in nucleus called chromatin, thread-
like stained (coloured) bodies seen in nucleus. The nucleosomes in nn
chromatin are seen as ‘beads-on-string’ structure when viewed under
electron microscope (EM) (Figure 5.4 b).
Theoretically, how many such beads (nucleosomes) do you imagine
83
are present in a mammalian cell?
The beads-on-string structure in chromatin is packaged to form
chromatin fibers that are further coiled and condensed at metaphase stage
of cell division to form chromosomes. The packaging of chromatin at higher
level requires additional set of proteins that collectively are referred to as
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E
Non-histone Chromosomal (NHC) proteins. In a typical nucleus, some
region of chromatin are loosely packed (and stains light) and are referred to
as euchromatin. The chromatin that is more densely packed and stains
dark are called as Heterochromatin. Euchromatin is said to be
transcriptionally active chromatin, whereas heterochromatin is inactive.
Transforming Principle
In 1928, Frederick Griffith, in a series of experiments with Streptococcus
pneumoniae (bacterium responsible for pneumonia), witnessed a
miraculous transformation in the bacteria. During the course of his
experiment, a living organism (bacteria) had changed in physical form.
When Streptococcus pneumoniae (pneumococcus) bacteria are grown
on a culture plate, some produce smooth shiny colonies (S) while others
produce rough colonies (R). This is because the S strain bacteria have a
mucous (polysaccharide) coat, while R strain does not. Mice infected with
the S strain (virulent) die from pneumonia infection but mice infected
with the R strain do not develop pneumonia.
84
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Conclusion
He concluded that the R strain bacteria had somehow been
transformed by the heat-killed S strain bacteria. Some ‘transforming
Transforming
principle’, transferred from the heat-killed S strain, had enabled the
R strain to synthesise a smooth polysaccharide coat and become virulent.
This must be due to the transfer of the genetic material. However, the
biochemical nature of genetic material was not defined from his
experiments. principle.
Biochemical Characterisation of Transforming Principle
Prior to the work of Oswald Avery, Colin MacLeod and Maclyn McCarty
(1933-44), the genetic material was thought to be a protein. They worked
I
to determine the biochemical nature of ‘transforming principle’ in Griffith's
experiment.
They purified biochemicals (proteins, DNA, RNA, etc.) from the
heat-killed S cells to see which ones could transform live R cells into
S cells. They discovered that DNA alone from S bacteria caused R bacteria
-
to become transformed.
They also discovered that protein-digesting enzymes (proteases) and
-
-M1933-44-proteinasthat
-
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supunatant
-
(Brit
Pellet
(E.(01i)
5
/Snake)
Alelet
86
5.2.2 Properties of Genetic Material (DNA versus RNA)
From the foregoing discussion, it is clear that the debate between proteins
versus DNA as the genetic material was unequivocally resolved from
Hershey-Chase experiment. It became an established fact that it is DNA
that acts as genetic material. However, it subsequently became clear that
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in some viruses, RNA is the genetic material (for example, Tobacco Mosaic
viruses, QB bacteriophage, etc.). Answer to some of the questions such as, RNA
why DNA is the predominant genetic material, whereas RNA performs
dynamic functions of messenger and adapter has to be found from the Its stable
differences between chemical structures of the two nucleic acid molecules. -
do more tune
Can you recall the two chemical differences between DNA and RNA?
↓
A molecule that can act as a genetic material must fulfill the following
criteria: -
Str Role-Ribosome
Catalytic Ribozyme
-
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Storage NA genetic material, but DNA being more stable is preferred for storage of
-
RNA was the first genetic material. There is now enough evidence to
suggest that essential life processes (such as metabolism, translation,
splicing, etc.), evolved around RNA. RNA used to act as
a genetic material as well as a catalyst (there are some
important biochemical reactions in living systems that
are catalysed by RNA catalysts and not by protein
enzymes). But, RNA being a catalyst was reactive and
hence unstable. Therefore, DNA--
has evolved from RNA
with chemical modifications that make it more stable.
-
DNA being double stranded and having complementary
-
strand further resists changes by evolving a process of
e
repair.
--
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and human cells. Matthew Meselson and Franklin Stahl performed the
following experiment in 1958:
-Scot Radioactive
2sotoope
(i) They grew E. coli in a medium containing 15 4 15
NH Cl ( N is the heavy
isotope of nitrogen) as the only nitrogen source for many
generations. The result was that 15N was incorporated into newly is
si
-
=
synthesised DNA (as well as other nitrogen >
containing compounds).
This heavy DNA molecule could be distinguished from the normal
-
DNA by centrifugation in a cesium chloride (CsCl) density gradient
(Please note that 15N is not a radioactive isotope, and it can be
gid
(ii) Then they transferred the cells into a medium with normal
G1 100
14
NH4Cl and took samples at various definite time intervals as
-
=
45-
independently on CsCl gradients to measure the densities of
DNA (Figure 5.7). 14
"So,
ay Can you recall what centrifugal force is, and think why a
molecule with higher mass/density would sediment faster?
The results are shown in Figure 5.7.
50%
hybrid
G1
az
=>
-
-
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Eukaryot
The experiments proved that the DNA in chromosomes also replicate
canosomal
2
semiconservatively.
Topoisomeran
-
Heliease-Unwinding
-2nd
into mutations. Furthermore, energetically replication is a very expensive
⑧
enzyme
process. Deoxyribonucleoside triphosphates serve dual purposes. In
addition to acting as substrates, they provide energy for polymerisation
brk H-Bord reaction (the two terminal phosphates in a deoxynucleoside triphosphates
-
are high-energy phosphates, same as in case of ATP).
In addition to DNA-dependent DNA polymerases, many additional
enzymes are required to complete the process of replication with high
degree of accuracy. For long DNA molecules, since the two strands of
lot of
errggre
DNA cannot be separated in its entire length (due to very high energy
requirement), the replication occur within a small opening of the DNA
helix, referred to as replication fork. The DNA-dependent DNA
polymerases catalyse polymerisation only in one direction, that is 5' 3' .
This creates some additional complications at the replicating fork.
Consequently, on one strand (the template with polarity 3' 5' ), the
replication is continuous, while on the other (the template with
90
-
DNA-DNA DNA
dependent polymeras
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MOLECULAR BASIS OF INHERITANCE
dig laggand
chromosomal anomaly). You will learn the detailed
nature of origin and the processes occurring at this
site, in higher classes.
only
formes-tRNA)
would be transcribed. 1
Why both the strands are not copied during transcription has the
simple answer. First, if both strands act as a template, they would code ↳
gene
PartyDNAwhichene
-
for RNA molecule with different sequences (Remember complementarity
does not mean identical), and in turn, if they code for proteins, the sequence
·
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define a gene in terms of DNA sequence. The DNA sequence coding for
-
tRNA or rRNA molecule also define a gene. However by defining a cistron
- -
Polycistro
- -
Monoulstry
-
only
as a segment of DNA coding for a polypeptide, the structural gene in a I
-DN A
nving
-
/tspitgene- only
definition of a gene in terms of a DNA segment. sukart
-
- -
Prokaryot
-
Inheritance of a character is also affected by promoter and regulatory
sequences of a structural gene. Hence, sometime the regulatory sequences
are loosely defined as regulatory genes, even though these sequences do
not code for any RNA or protein.
-
structural
-
tRNA brings aminoacids and reads the genetic code, and rRNAs playI
and catalytic role during translation. There is single
-
DNA-dependent RNA polymerase that catalyses transcription of all types
of RNA in bacteria. RNA polymerase binds to promoter and initiates
->
transcription (Initiation). It uses nucleoside triphosphates as substrate
breakage of thebond
-
Here
Helicat
not down
by
cofactor. Rather RNA
pol does
itself
93
Lu ⑧
e
Par
peptidyltranet
is
Figure 5.10 Process of Transcription in Bacteria
en
referent
=23r Addy
-
EU E MessengenateRiboso
-
e
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E enzyme. Once the polymerases reaches the terminator region, the nascent
RNA falls off, so also the RNA polymerase. This results in termination of
transcription.
An intriguing question is that how is the RNA polymerases able
to catalyse all the three steps, which are initiation, elongation and
termination. The RNA polymerase is only capable of catalysing the
Signa
process of elongation. It associates transiently with initiation-factor (σ)
Rho and termination-factor (ρ) to initiate and terminate the transcription,
-> respectively. Association with these factors alter the specificity of the
RNA polymerase to either initiate or terminate (Figure 5.10).
In bacteria, since the mRNA does not require any processing to become
active, and also since transcription and translation take place in the same
compartment (there is no separation of cytosol and nucleus in bacteria),
a
many times the translation can begin much before the mRNA is fully
transcribed. Consequently, the transcription and translation can be coupled
in bacteria.
In eukaryotes, there are two additional complexities –
inypotayotout
-
(i) There are at least three RNA polymerases in the nucleus (in addition
in
-
to the RNA polymerase found in the organelles). There is a clear
-
cut division of labour. The RNA polymerase I transcribes rRNAs
- -
sty
FreI
- i
-
I
M -> (200-300
94
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MOLECULAR BASIS OF INHERITANCE
Lud (28S, 18S, and 5.8S), whereas the RNA polymerase III is responsible
- -
Im nucleus).
Z
-
-
In
e
Trangppti
Post
e
-
(ii) The second complexity is that the primary transcripts contain both
the exons and the introns and are non-functional. Hence, it is
E subjected to a process called splicing where the introns are removed
and exons are joined in a defined order. 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 (Figure 5.11).
The significance of such complexities is now beginning to be
understood. The split-gene arrangements represent probably an ancient
feature of the genome. The presence of introns is reminiscent of antiquity,
and the process of splicing represents the dominance of RNA-world. In
recent times, the understanding of RNA and RNA-dependent processes
in the living system have assumed more importance.
Bases
transfer of genetic information from a polymer of nucleotides to synthesise
a polymer of amino acids. Neither does any complementarity exist between Total=64 Codous.
nucleotides and amino acids, nor could any be drawn theoretically. There
existed ample evidences, though, to support the notion that change in
nucleic acids (genetic material) were responsible for change in amino acids
in proteins. This led to the proposition of a genetic code that could direct
the sequence of amino acids during synthesis of proteins.
If determining the biochemical nature of genetic material and the
structure of DNA was very exciting, the proposition and deciphering of
genetic code were most challenging. In a very true sense, it required
involvement of scientists from several disciplines – physicists, organic
chemists, biochemists and geneticists. It was George Gamow, a physicist,
who argued that since there are only 4 bases and if they have to code for
20 amino acids, the code should constitute a combination of bases. He 95
suggested that in order to code for all the 20 amino acids, the code should
be made up of three nucleotides. This was a very bold proposition, because
a permutation combination of 43 (4 × 4 × 4) would generate 64 codons;
generating many more codons than required.
Providing proof that the codon was a triplet, was a more daunting
task. The chemical method developed by Har Gobind Khorana was
=>
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can from
DA
I Severo Ochoa enzyme (polynucleotide phosphorylase) was also helpful
in polymerising RNA with defined sequences in a template independent
-
I
without manner (enzymatic synthesis of RNA). Finally a checker-board for genetic
-
Gudependent
-
. DNA Table 5.1: The Codons for the Various Amino Acids ⑤
polymera
RNA
Phenyle
Nirenberg only homopoly
mel
-
karGobinda-Homopoly me
i
co-polycal
codp"."Synously
(ii) Some amino acids are coded by more than one codon, hence
tophan geti
>
UUU would code for Phenylalanine (phe). Some exceptions to this
- =
rule have been found in mitochondrial codons, and in some
protozoans. thuwic UGAStoPoy
=
(v) AUG has dual functions. It codes for Methionine (met) , and it
- -
only, code
sequence of amino acid coded by it (take help of the checkerboard):
-AUG UUU UUC UUC UUU UUU UUC-
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MOLECULAR BASIS OF INHERITANCE
Now try the opposite. Following is the sequence of amino acids coded
by an mRNA. Predict the nucleotide sequence in the RNA:
Met-Phe-Phe-Phe-Phe-Phe-Phe
Do you face any difficulty in predicting the opposite?
Can you now correlate which two properties of genetic code you have
learnt?
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complementary to
the code, and it also
has an amino acid
u acceptor end to
which it binds to
-
amino acids.
tRNAs are specific
for each amino acid
(Figure 5.12). For
Figure 5.12 tRNA - the adapter molecule initiation, there is
another specific tRNA that is referred to as initiator tRNA. There are no
-
tRNAs for stop codons. In figure 5.12, the secondary structure of tRNA
has been depicted that looks like a clover-leaf. In actual structure, the
tRNA is a compact molecule which looks like inverted L.
5.7 TRANSLATION
Translation refers to the process of polymerisation of amino acids to
form a polypeptide (Figure 5.13). 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
98 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
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MOLECULAR BASIS OF INHERITANCE
43
[ =
datanee
- >
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
-
matter
the tRNA anticodon. The ribosome moves from codon to codon along the
mRNA. Amino acids are added one by one, translated into Polypeptide
t 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.
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only prokaryot
5.8.1 The Lac operon
The elucidation of the lac operon was also a result of a close association
between a geneticist, Francois Jacob and a biochemist, Jacque Monod. They
-
were the first to elucidate a transcriptionally regulated system. In lac operon
diaanhibitor
(here lac refers to lactose), a polycistronic structural gene is regulated by a
common promoter and regulatory genes. Such arrangement is very common
in bacteria and is referred to as operon. To name few such examples, lac
operon, trp operon, ara operon, his operon, val operon, etc.
gene
↓
The lac operon consists of one regulatory gene (the i gene – here the
=--
forms
Represstein
term i does not refer to inducer, rather it is derived from the word inhibitor)
- -
and three structural genes (z, y, and a). The i gene codes for the repressor
--
of the lac operon. The z gene codes for beta-galactosidase (β-gal), which
--
is primarily responsible for the hydrolysis of the disaccharide, lactose
100 into its monomeric units, galactose and glucose. The y gene codes for
-
as well, the genes present in the operon are needed together to function
③
promote gene
in the same or related metabolic pathway (Figure 5.14).
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(lactose
->
Polycistic
Clactose bind
about allow itto
operator
Figure 5.14 The lac Operon
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Goals of HGP
Some of the important goals of HGP were as follows:
(i) Identify all the approximately 20,000-25,000 genes in human DNA;
(ii) Determine the sequences of the 3 billion chemical base pairs that
make up human DNA;
(iiii) Store this information in databases;
(iv) Improve tools for data analysis;
(v) Transfer related technologies to other sectors, such as industries;
(vi) Address the ethical, legal, and social issues (ELSI) that may arise
from the project.
The Human Genome Project was a 13-year project coordinated by
102 the U.S. Department of Energy and the National Institute of Health. During
the early years of the HGP, the Wellcome Trust (U.K.) became a major
partner; additional contributions came from Japan, France, Germany,
China and others. The project was completed in 2003. Knowledge about
the effects of DNA variations among individuals can lead to revolutionary
new ways to diagnose, treat and someday prevent the thousands of
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MOLECULAR BASIS OF INHERITANCE
⑳ 0-
Sequence Annotation). For sequencing, the total DNA from a cell is
isolated and converted into random fragments of relatively smaller sizes
- - >
(recall DNA is a very long polymer, and there are technical limitations in nost-E.coli
BA), YAC.
sequencing very long pieces of DNA) and cloned in suitable host using
-
chromosomes).
The fragments were sequenced using automated DNA sequencers that
worked on the principle of a method developed by Frederick Sanger.
(Remember, Sanger is also credited for developing method for
determination of amino acid
sequences in proteins). These
sequences were then arranged based
on some overlapping regions
present in them. This required
generation of overlapping fragments
for sequencing. Alignment of these
sequences was humanly not
possible. Therefore, specialised
computer based programs were
developed (Figure 5.15). These
sequences were subsequently
annotated and were assigned to each
103
chromosome. The sequence of
chromosome 1 was completed only
in May 2006 (this was the last of the
24 human chromosomes – 22 Figure 5.15 A representative diagram of human
autosomes and X and Y – to be genome project
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mux- (ii) The average gene consists of 3000 bases, but sizes vary greatly, with
the largest known human gene being dystrophin at 2.4 million bases.
Testis genes.
determining th 95Junk.
tag
(v) Less than 2 per cent of the genome codes for proteins.
(vi) Repeated sequences make up very large portion of the human genome.
(vii) Repetitive sequences are stretches of DNA sequences that are
repeated many times, sometimes hundred to thousand times. They
are thought to have no direct coding functions, but they shed light
malte
on chromosome structure, dynamics and evolution.
(viii) Chromosome 1 has most genes (2968), and the Y has the fewest (231).
E
Langer
(ix) Scientists have identified about 1.4 million locations where single-
- >
base DNA differences (SNPs – single nucleotide polymorphism,
-
pronounced as ‘snips’) occur in humans. This information promises
to revolutionise the processes of finding chromosomal locations for
disease-associated sequences and tracing human history.
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MOLECULAR BASIS OF INHERITANCE
broader scale. They can study all the genes in a genome, for example, all
- -
(Motsam
two individuals.
DNA fingerprinting involves identifying differences in some specific
regions in DNA sequence called as repetitive DNA, because in these
sequences, a small stretch of DNA is repeated many times. These repetitive
DNA as
gat
atmnist af
DNA are separated from bulk genomic DNA as different peaks during
my
density gradient centrifugation. The bulk DNA forms a major peak and sitifive
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
e
genome. These sequence show high degree of polymorphism and form
equNTR
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
Monozygotic
only twin same
have
of paternity testing, in case of disputes.
As polymorphism in DNA sequence is the basis of genetic mapping
UnitR.
of human genome as well as of DNA fingerprinting, it is essential that we
understand what DNA polymorphism means in simple terms.
Polymorphism (variation at genetic level) arises due to mutations. (Recall
different kind of mutations and their effects that you have already
studied in Chapter 4, and in the preceding sections in this chapter.) 105
New mutations may arise in an individual either in somatic cells or in
the germ cells (cells that generate gametes in sexually reproducing
organisms). If a germ cell mutation does not seriously impair individual’s
ability to have offspring who can transmit the mutation, it can spread to
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I
(i) isolation of DNA,
order (ii) digestion of DNA by restriction endonucleases,
Lu Zu
(iii) separation of DNA fragments by electrophoresis,
(iv) transferring (blotting) of separated DNA fragments to synthetic
(Southern Blotting)
membranes, such as nitrocellulose or nylon,
IV-b-make it 5S
(v) hybridisation using labelled VNTR probe, and
=>
Radioactive.
-
artificial
representation of DNA fingerprinting is shown in Figure 5.16.
The VNTR belongs to a class of satellite DNA referred to as mini-satellite.
Complementin
-
A small DNA sequence is arranged tandemly in many copy numbers. The
copy number varies from chromosome to chromosome in an individual.
The numbers of repeat show very high degree of polymorphism. As a
result the size of VNTR varies in size from 0.1 to 20 kb. Consequently,
=
after hybridisation with VNTR probe, the autoradiogram gives many bands
of differing sizes. These bands give a characteristic pattern for an individual
106 DNA (Figure 5.16). It differs from individual to individual in a population
stioy except in the case of monozygotic (identical) twins. The sensitivity of the
exof
technique has been increased by use of polymerase chain reaction (PCR–
you will study about it in Chapter 9). Consequently, DNA from a single
-
cell is enough to perform DNA fingerprinting analysis. In addition to
-
application in forensic science, it has much wider application, such as
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107
in determining population and genetic diversities. Currently, many
different probes are used to generate DNA fingerprints.
-
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SUMMARY
Nucleic acids are long polymers of nucleotides. While DNA stores genetic
information, RNA mostly helps in transfer and expression of information.
Though DNA and RNA both function as genetic material, but DNA being
chemically and structurally more stable is a better genetic material.
However, RNA is the first to evolve and DNA was derived from RNA. The
hallmark of the double stranded helical structure of DNA is the hydrogen
bonding between the bases from opposite strands. The rule is that
Adenine pairs with Thymine through two H-bonds, and Guanine with
Cytosine through three H-bonds. This makes one strand
complementary to the other. The DNA replicates semiconservatively,
the process is guided by the complementary H-bonding. A segment of
DNA that codes for RNA may in a simplistic term can be referred as
gene. During transcription also, one of the strands of DNA acts a
template to direct the synthesis of complementary RNA. In bacteria,
the transcribed mRNA is functional, hence can directly be translated.
In eukaryotes, the gene is split. The coding sequences, exons, are
interrupted by non-coding sequences, introns. Introns are removed
and exons are joined to produce functional RNA by splicing. The
messenger RNA contains the base sequences that are read in a
combination of three (to make triplet genetic code) to code for an amino
acid. The genetic code is read again on the principle of complementarity
by tRNA that acts as an adapter molecule. There are specific tRNAs for
every amino acid. The tRNA binds to specific amino acid at one end
and pairs through H-bonding with codes on mRNA through its
anticodons. The site of translation (protein synthesis) is ribosomes,
which bind to mRNA and provide platform for joining of amino acids.
One of the rRNA acts as a catalyst for peptide bond formation, which is
an example of RNA enzyme (ribozyme). Translation is a process that
has evolved around RNA, indicating that life began around RNA. Since,
transcription and translation are energetically very expensive
processes, these have to be tightly regulated. Regulation of transcription
is the primary step for regulation of gene expression. In bacteria, more
than one gene is arranged together and regulated in units called as
operons. Lac operon is the prototype operon in bacteria, which codes
for genes responsible for metabolism of lactose. The operon is regulated
by the amount of lactose in the medium where the bacteria are grown.
Therefore, this regulation can also be viewed as regulation of enzyme
synthesis by its substrate.
Human genome project was a mega project that aimed to sequence
every base in human genome. This project has yielded much new
108 information. Many new areas and avenues have opened up as a
consequence of the project. DNA Fingerprinting is a technique to find
out variations in individuals of a population at DNA level. It works on
the principle of polymorphism in DNA sequences. It has immense
applications in the field of forensic science, genetic biodiversity and
evolutionary biology.
Rationalised 2023-24
MOLECULAR BASIS OF INHERITANCE
EXERCISES
1 Group the following as nitrogenous bases and nucleosides:
Adenine, Cytidine, Thymine, Guanosine, Uracil and Cytosine.
2. If a double stranded DNA has 20 per cent of cytosine, calculate the per
cent of adenine in the DNA.
3. If the sequence of one strand of DNA is written as follows:
5' -ATGCATGCATGCATGCATGCATGCATGC-3'
Write down the sequence of complementary strand in 5' →3' direction.
4. If the sequence of the coding strand in a transcription unit is written
as follows:
5' -ATGCATGCATGCATGCATGCATGCATGC-3'
Write down the sequence of mRNA.
5. Which property of DNA double helix led Watson and Crick to hypothesise
semi-conservative mode of DNA replication? Explain.
6. Depending upon the chemical nature of the template (DNA or RNA)
and the nature of nucleic acids synthesised from it (DNA or RNA), list
the types of nucleic acid polymerases.
7. How did Hershey and Chase differentiate between DNA and protein in
their experiment while proving that DNA is the genetic material?
8. Differentiate between the followings:
(a) Repetitive DNA and Satellite DNA
(b) mRNA and tRNA
(c) Template strand and Coding strand
9. List two essential roles of ribosome during translation.
10. In the medium where E. coli was growing, lactose was added, which
induced the lac operon. Then, why does lac operon shut down some
time after addition of lactose in the medium?
11. Explain (in one or two lines) the function of the followings:
(a) Promoter
(b) tRNA
(c) Exons
12. Why is the Human Genome project called a mega project?
13. What is DNA fingerprinting? Mention its application.
14. Briefly describe the following:
(a) Transcription 109
(b) Polymorphism
(c) Translation
(d) Bioinformatics
Rationalised 2023-24