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4. SOME IMPORTANT QUESTION - ANSWER FOR IGNOU BIOCHEMISTRY [CHE 9]
NUCLEIC ACIDS
Q. 1. What are the principal constituents of nucleic acids? Mention the differences
between a nucleoside and a nucleotide. Draw the structure of ATP.[Nucleic acids क
े मु य
अवयव या होते ह? Nucleoside और nucleotide म अंतर बताय I ATP क संरचना बनाएं ]
Ans. Nucleic acids are polymerized chains of nucleotides (polynucleotides) joined by
phosphodiester bonds. The two types of nucleic acids are deoxyribonucleic acid (DNA) and
ribonucleic acids (RNA).
Deoxyribonucleic acid (DNA) - is an universal genetic material and a fundamental unit of
heredity. DNA consists of two independent polydeoxyribonucleotide strands in the form of a right
handed double helix inside the cell. The backbone of DNA is made of a chain of deoxyribose
sugar units joined together by phosphodiester bonds (between 3' end of one sugar and the 5'
end of the next sugar). In addition, four types of nitrogeous bases [two purines - adenine (A) &
guanine (G) and two pyrimidines - cytosine (C ) & thymine (T)] are also attached to sugar units
and projected on inside of the backbone. The amount of A is equal to T, whereas the amount of
C is equal to G (A=T; C=G). The bases of one strand are hydrogen bonded to the bases of the
other strand of the double helix. The base pairing is always between adenine and thymine by
two hydrogen bonds and between cytosine and guanine by three hydrogen bonds. In addition to
these hydrogen bonds, there are some interactions like hydrophobic or stacking interactions
which provide further stability to the double helix.
Ribonucleic acids (RNA) - are single stranded polymers of ribonucleotides in which the pentose
sugar is ribose instead of deoxyribose which is present in DNA. The ribose sugar units are
joined together by phosphodiester bonds to form a strand to which four types of nitrogenous
bases (adenine, guanine, cytosine and uracil) are attached. In RNA uracil nitrogenous base is
present instead of thymine which is present in DNA. RNAs are generally single stranded except
a few viral RNA molecules which are double stranded. The RNAs are the "middle man" of
information transfer and there are three main types of RNAs based on the different roles played
by them in the biosynthesis of proteins and polypeptides. These are ribosomal RNA (rRNA),
messenger RNA (mRNA) and transfer RNA (tRNA).
[Nucleic acids phodphodiester bonds वारा जुड़े nucleotides क polymerized chains होते ह अथात
nucleotides क
े polymer होते ह I दो कार क
े nucleic acids ह - deoxyribonucleic acid (DNA) और
ribonucleic acids (RNA) I
Deoxyribonucleic acid (DNA) एक universal genetic material और आनुवं शकता (heredity) क मौ लक
इकाई (fundamental unit) है I DNA molecule म दो वतं (independent) polydeoxyribonucleotide
strands होते ह जो को शका क
े अंदर एक right handed double helix क
े प म होते ह I DNA molecule क
backbone phosphodiester bonds वारा जुड़े हुए deoxyribose sugar units क एक ृंखला से बनी होती है
और इसक
े अ त र त इसम nitrogenous bases [two purines - adenine (A) & guanine (G) and two
pyrimidines - cytosine (C ) & thymine (T)] भी जुड़े होते ह जो इस backbone से अंदर क ओर projected
होते ह I A क मा ा T क
े बराबर और C क मा ा G क
े बराबर होती है (A=T; C=G) I एक strand क
े bases दूसरे
strand क
े bases से hydrogen bonds वारा जुड़े होते ह और यह base pairing हमेशा adenine (A) एवं
thymine (T) क
े बीच दो hydrogen bonds वारा और cytosine (C) एवं guanine (G) क
े बीच तीन
2
hydrogen bonds वारा होती है I इसक
े अ त र त क
ु छ अ य interactions जैसे hydrophobic और stacking
interactions भी होते ह जो double helix को ि थरता दान करते ह I]
Ribonucleic acid (RNA) - ये nucleotides क
े single stranded polymer होते ह और इनम शुगर units
ribose होती ह (DNA म deoxyribose sugar units होती ह) I DNA molecule क तरह ह RNA क
backbone phosphodiester bonds वारा जुड़े हुए ribose sugar units क एक ृंखला से बनी होती है और
इसक
े अ त र त इसम nitrogenous bases [दो purines - adenine (A) एवं guanine (G) and दो
pyrimidines - cytosine (C) एवं uracil (U)] भी जुड़े होते ह I RNAs सामा य तौर पर single stranded होते
ह क तु क
ु छ virus म ये double stranded होते ह I RNAs information transfer म बचौ लए (middle
man) क भू मका नभाते ह I proteins और polypeptides क biosynthesis म अपनी -अपनी भू मका क
े
आधार पर RNA तीन कार क
े होते ह - ribosomal RNA (rRNA), messenger RNA (mRNA) and transfer
RNA (tRNA)]
Difference between nucleoside and nucleotide:
1. A nucleoside consists of a sugar and a nitrogeous base without the phosphate group
whereas a nucleotide consists of a phosphate group, a sugar and a nitrogenous base.
[एक nucleoside एक sugar molecule और एक nitrogenous base से मलकर बना होता है और
इसम कोई phosphate group नह ं होता है जब क nucleotide म sugar, nitrogenous base और एक
phosphate group होता है]
2. The bond formed in a nucleoside is between C1 of the pentose sugar and either N1 of
pyrimidine base or N9 of the purine base whereas in nucleotide a phosphate group is
attached to C5 of the pentose sugar of the nucleoside by a phosphodiester linkage.
[nucleoside म pentose sugar क
े C1 और pyrimidine base क
े N1 या purine base क
े N9 क
े बीच
एक bond बना होता है जब क nucleotide म nucleoside क
े pentose sugar क
े C5 से एक
phosphate group भी एक phodphodiester bond वारा जुड़ा होता है]
Nucleosides Nucleotide
3
Structure of ATP
Q. 2. Describe the salient features of Watson and Crick model of DNA. What are the
important functions of DNA?[DNA क
े Watson और Crick model क मु य वशेषताएं लख I DNA
क
े मु य काय लख ]
Ans. Salient features of Watson and Crick model od DNA :
● DNA molecule has two independent polydeoxyribonucleotide strands which are in the
form of a right handed double helix inside the cell.
[DNA molecule म दो वतं (independent) polydeoxyribonucleotide strands होते ह जो
को शका क
े अंदर एक right handed double helix क
े प म होते ह I]
● The two strands in DNA double helix are antiparallel, which means that the 5' end of one
strand is paired with the 3' end of its complementary strand (and vice versa).
[DNA क
े double helix क
े दोन strands antiparallel होते ह, अथात एक strand का 5' छोर दूसरे
complementary strand क
े 3' छोर क
े साथ जोड़ी बनाता है ]
● The DNA backbone consists of a chain of deoxyribose sugar units joined by 3',
5'-phosphodiester bonds (between 3' end of one sugar and the 5' end of the next sugar)
and these are projected outside of the backbone.
[DNA molecule क backbone 3', 5'-phosphodiester bonds वारा जुड़े हुए deoxyribose sugar
units क एक ृंखला से बनी होती है (between 3' end of one sugar and the 5' end of the next
sugar) जो इस backbone क
े बाहर क तरफ projected रहते ह ]
● In addition, two purine (adenine, guanine) and two pyrimidine (thymine, cytosine) bases
are also attached to this backbone and projected inside of the backbone.
[इसक
े अ त र त इसम nitrogenous bases (two purines - adenine & guanine and two
pyrimidines - cytosine & thymine) भी जुड़े होते ह जो इस backbone से अंदर क ओर projected
होते ह]
● The bases of one strand are attached to the bases of the other strand by hydrogen
bonds and this base pairing is always by two hydrogen bonds between adenine (A) &
thymine (T) and by three hydrogen bonds between cytosine(C) & guanine (G).
[एक strand क
े bases दूसरे strand क
े bases से hydrogen bonds वारा जुड़े होते ह और यह base
pairing हमेशा adenine (A) एवं thymine (T) क
े बीच दो hydrogen bonds वारा और cytosine(C)
एवं guanine (G) क
े बीच तीन hydrogen bonds वारा होती है ]
● A small groove, called the minor groove (1.2 nm), and a deep groove, known as the
major groove (2.2 nm), run across the DNA helix which are created by the offset pairing
of the two strands.
[DNA क
े दोन strands क ऑफसेट जोड़ी क
े कारण पूरे DNA helix म small groove (1.2 nm) और
deep groove (2.2 nm) होते ह ]
4
Functions of DNA -
● DNA is an universal genetic material and a fundamental unit of heredity. It is responsible
for the inheritance and transmission of specific characteristics from one generation to the
other.
[DNA universal genetic material और आनुवं शकता (heredity) क मौ लक इकाई (fundamental
unit) है I यह वंशानु म और एक पीढ़ से दूसर पीढ़ म व श ट वशेषताओं क
े ह तांतरण क
े लए
िज़ मेदार होता है]
● DNA dictates how a human or animal develops and reproduces, and eventually dies.
[DNA तय करता है क एक human या animal क
ै से वक सत, जनन करता है और अंत म क
ै से मृ यु
को ा त होता है]
● DNA stores instructions for making other large molecules, called proteins.
[DNA अ य बड़े molecules proteins क
े नमाण क
े लए वां छत नदश का सं ह करता है]
● DNA is also involved in [DNA इनम भी शा मल होता है] :
(i) Equal distribution of DNA during the cell division [cell division क
े समय DNA क
े समान
वतरण म ]
(ii) Mutations i.e.The changes which occur in the DNA sequences [DNA sequence म होने
वाले प रवतन म ]
(iii) Transcription (RNA synthesis) [RNA क
े सं लेषण (transcription) म ]
(iv) Cellular Metabolism [cells क
े अंदर होने वाल metabolism म ]
(v) DNA fingerprinting [DNA fingerprinting म ]
(vi) Gene Therapy [gene therapy म]
Q. 3. What are the different types of RNA? Describe briefly.[ व भ न कार क
े RNA या होते ह,
सं ेप म वणन कर I]
Ans. RNAs have been classified into three types viz. ribosomal RNA (rRNA), messenger RNA
(mRNA) and transfer RNA (tRNA), depending on the basis of their different roles in biosynthesis
of proteins and polypeptides.
Ribosomal RNA - This constitutes about 66% of total RNA present in the ribosomes of a cell. It
is synthesized in the nucleolus of the nucleus. These are non coding RNAs and are single
stranded. The G:C and A:U ratios are unequal. In E. coli three forms of rRNA (5S, 16S, 23S) are
found which differ in their base ratios and sequences. Likewise, in eukaryotic cells 5S, 7S, 18S
and 28S forms of rRNAs are found. The formation of hydrogen bonds between base pairs
results in the formation of the hairpin type turns in the rRNA molecules as found in tRNA. The
precise role of rRNA is not clear but they are sites of polypeptide synthesis and they form
complex with mRNA. The rRNA ensures the proper alignment of the mRNA, tRNA and the
ribosomes.
Messenger RNA - These are so called because they carry the 'message' from the coded DNA
and execute/guide the synthesis of polypeptides. mRNAs are much shorter than rRNA and have
5
triplets of bases (codons) complementary to the DNA and each codon is the code for a
particular amino acid. The sequence of these codons on the mRNA guides the synthesis of the
particular polypeptide. The length of these mRNAs varies depending upon the lengths of
polypeptides for which they are coded. mRNA is synthesized from DNA in the nucleus by a
process called 'transcription' in which the genetic code (sequence of bases) is transcribed from
one strand of the DNA molecule in the form of a single strand of mRNA with the help of an
enzyme called RNA polymerase. After transcription, the mRNA passes into the cytoplasm and
then to ribosomes where it serves as the template for the process of translation during protein
synthesis.
Transfer RNA - These are small molecules with 60-95 nucleotides. These are single stranded
too but adopt a unique cloverleaf like structure due to extensive complementary base pairing of
adenine:uracil and guanine:cytosine. Their function is to carry the individual amino acids to the
site of protein synthesis and for this purpose they carry a complementary triplet of bases called
the 'anticodon' which recognizes the codon of mRNA for a particular amino acid, thus acting as
an interpreter in the process of translation in protein synthesis. The 3'- end ribose unit of tRNA
has an OH group which can attach a particular amino acid by an ester bond forming
aminoacyl-tRNA (charged form of tRNA). Each of the 20 amino acids found in proteins has at
least one corresponding tRNA. The tRNA sometimes has unusual or minor bases like inosine,
pseudouridine etc. The exact role of these unusual bases is not clearly known but these
somehow protect the tRNA degradation by enzyme RNAase. Also these are involved in
stabilization of the tertiary structure of tRNA.
[RNA को protein और polypeptide synthesis म उनक भू मका क
े आधार पर तीन कार म वग कृ त कया
गया है - ribosomal RNA (rRNA), messenger RNA (mRNA) और transfer RNA (tRNA) I
Ribosomal RNA - यह एक cell क
े ribosomes म उपि थत क
ु ल RNA का लगभग 66% होता है I इसका
सं लेषण nucleus क
े nucleolus म होता है I ये non-coding RNA होते ह और एक strand वाले होते ह I इनम
G:C और A:U क
े अनुपात असमान होते ह I E. coli म rRNA क
े तीन प (5S, 16S, 23S) होते ह और इनम
base ratios और sequences म भ नता होती है I इसी कार, eukaryotic cells म rRNA क
े चार प (5S,
7S, 18S और 28S) पाए जाते ह I rRNA क
े base pairs क
े बीच hydrogen bonds बनने क
े कारण एक
hairpin type का प बन जाता है, जैसा tRNA म भी होता है I rRNA क ब क
ु ल प ट भू मका क
े बारे म
प टता नह ं है क तु ये polypeptide synthesis क
े काय थल होते ह और ये mRNA क
े साथ complex बनाते
ह I mRNA, tRNA और ribosomes क
े बीच संरेखण (alignment) को rRNA सु नि चत करते ह I
Messenger RNA - इ ह messenger RNA इस लए कहते ह य क ये DNA क
े coded message को carry
करने और polypeptide synthesis को न पा दत करने म भू मका नभाते ह I ये rRNA से काफ़ छोटे होते ह
और इनम DNA क
े complementary bases क
े triplets (codons) होते ह और येक codon एक व श ट
amino acid का code होता है I mRNA पर इन codons का जो sequence होता है वो एक नि चत
polypeptide क synthesis का माग दशन करता है I इन mRNA क ल बाई उन polypeptides क ल बाई पर
नभर करती है िजनक
े लए ये mRNA coded होते ह I mRNA क synthesis nucleus म DNA से होती है और
इस या को 'transcription' कहते ह िजसम DNA क
े एक strand से genetic code (sequence of bases)
को एक mRNA क
े एक single strand क
े प म अनुले खत (transcribe) कया जाता है, एक enzyme RNA
polymerase क सहायता से I अनुलेखन (transcription) क
े बाद यह mRNA cytoplasm म पहुँचता है और
फर ribosomes पर पहुँचता है जहाँ यह protein synthesis क translation या म एक template क
भू मका नभाता है I
Transfer RNA - ये छोटे molecules होते ह िजनम 60-95 nucleotides होते ह I ये भी single stranded होते
ह क तु adenine:uracil और guanine:cytosine क अ य धक base pairing क
े कारण ये अनूठा cloverleaf
6
जैसी संरचना बना लेते ह I इनका काय होता है वैयि तक amino acids को protein synthesis क site पर ले
जाना और इसक
े लए इनम एक complementary bases का एक triplet होता है िजसे anticodon कहते ह I ये
anticodon mRNA क
े एक नि चत amino acid क
े codon को पहचान कर protein synthesis क
translation या म एक दुभा षया (interpreter) क भू मका नभाता है I tRNA क
े 3'- end क
े ribose unit क
े
OH group से एक नि चत amino acid एक ester bond वारा जुड़ कर aminoacyl-tRNA (charged form
of tRNA) बनता है I proteins म पाए जाने वाले 20 amino acids म से येक amino acid का कम से कम
एक अनु प (corresponding) tRNA होता है I कभी-कभी tRNA म unusual या minor bases जैसे - inosine,
pseudouridine आ द भी पाए जाते ह I इन unusual bases क
े काय क
े बारे म प टता नह ं है क तु ये tRNA
क
े RNAase enzyme वारा कये जाने वाले अध:पतन (degradation) को कसी तरह रोकते ह I इसक
े
अ त र त ये tRNA क
े tertiary structure को ि थरता दान करने म भी भू मका नभाते ह I
Q. 4. What are unusual bases? Write the major differences between RNA and
DNA.[असामा य bases या होते ह? RNA और DNA म मु य अंतर लख I]
Ans.
Unusual bases - In addition to primary nucleobases, DNA and RNA also contain other
(non-primary) unusual bases that have been modified after the nucleic acid chain has been
formed. These are formed by enzymatic modifications of the existing polynucleotides. In DNA,
the most common modified bases are 5-methylcytosine (m5C) in animals and higher plants, N6
methyladenosine in bacterial DNA and 5-hydroxymethylcytosine in animals and bacteria
infected with certain bacteriophages. The minor or unusual bases found in RNA, especially in
tRNA are Inosine, 1-methylinosine, N2-dimethylguanosine, ribothymidine, dihydrouridine,
pseudouridine, 7-methylguanosine and 4-thiouridine. The exact role of these unusual bases is
not clearly known. These somehow protect the tRNA degradation by RNAase. Also these are
involved in stabilization of the tertiary structure of tRNA. In DNA these unusual bases somehow
play some role in regulating or protecting the genetic information.
[DNA और RNA म सामा य (primary) nucleobases क
े अ त र त क
ु छ असामा य
(unusual/non-primary) bases भी पाए जाते ह जो nucleic acid chain बनने क
े बाद modify होकर बनते ह
और ये पहले से मौजूद nucleotides क
े enzymatic modifications वारा बनते ह I DNA म आमतौर पाए जाने
वाले modified bases ह - 5-methylcytosine (m5C) animals और higher plants म , N6
methyladenosine bacterial DNA म और 5-hydroxymethylcytosine animals और bacteria म जो क
ु छ
bacteriophages से सं मत ह I RNA म आमतौर पर पाए जाने वाले unusual या असामा य bases,
वशेषतः tRNA म, ह - Inosine, 1-methylinosine, N2-dimethylguanosine, ribothymidine,
dihydrouridine, pseudouridine, 7-methylguanosine और 4-thiouridine I इन unusual bases क
े
नि चत काय क
े बारे म प ट जानकार नह ं है क तु ये tRNA क
े RNAase enzyme वारा कये जाने वाले
अध:पतन (degradation) को कसी तरह रोकते ह और tRNA क
े tertiary structure को ि थरता दान करने म
भी भू मका नभाते ह I DNA म ये unusual bases DNA क genetic information क
े नयं ण और संर ण म
क
ु छ भू मका नभाते ह ]
7
Difference between DNA and RNA -
● DNA contains deoxyribose sugar but RNA contains ribose sugar
[DNA म शुगर deoxyribose होती है और RNA म ribose sugar होती है ]
● DNA is a double stranded molecule but RNA is a single stranded molecule.
[DNA म दो strands होते ह जब क RNA म एक strand होता है ]
● DNA contains adenine, guanine, cytosine and thymine but RNA contains uracil in place
of thymine. [DNA म adenine, guanine, cytosine और thymine bases होते ह जब क RNA म
thymine क
े थान पर uracil base होता है ]
● DNA is self replicating but RNA is synthesized by transcription of DNA. [DNA वयं से
replicate होता है (self replicating) जब क RNA DNA क
े transcription से बनता है ]
● DNA contains genetic information for the cell but RNA uses the information stored in
DNA to make proteins. [DNA म genetic information न हत रहती है जब क RNA DNA क इस
information को protein synthesis म उपयोग करता है ]
Q. 5. What are the different forms of DNA? Mention the main differences between the B-
DNA and Z-DNA.[DNA क
े व भ न प या होते ह? B-DNA और Z-DNA म मु य अंतर या ह?]
Ans. It has been found that DNA can occur in different three dimensional forms other than
Watson and Crick DNA structure. However, these variations generally do not affect the key
properties of DNA defined by Watson and Crick such as strand complementarity, antiparallel
strands, and the requirement for A=T and G≡C base pairing but there may be slight adjustment
in the angle of rotation between the adjacent base pairs. The different forms of DNA molecules
are: A-DNA, B-DNA, Z-DNA, C-DNA, D-DNA, E-DNA
A-DNA - has 11 base pairs per helical turn which is more than the average. The double helix is
right handed but a little wider but bases lie tilted to the helix axis instead of perpendicular to the
helix The major groove is deeper but minor groove is shallower. This form appears at 75%
relative humidity and require the presence of sodium, potassium or caesium ions.
B-DNA - It is the famous Watson & Crick structure and is the most common conformation in the
living cells and is a right handed helix. It is the most stable structure under physiological
conditions. It has 10.5 base pairs per helical turn. It requires high humidity (92%) and solutions
of low ionic strength.
Z-DNA - It is a left-handed double helix with a zig-zag pattern in the sugar-phosphate backbone.
It has a very different structure when compared with A-DNA and B-DNA which are right-handed
double helix structures. It has 12 base pairs per helical turn. The major groove is barely
apparent and the minor groove is narrow and deep. The Z-DNA has been found in a large
number of living organisms including mammals, protozoans and several plant species.
C-DNA - It is also a double helical conformations of DNA but has 9.33 base pairs per helical
turn. The C-form DNA is not very stable and does not occur naturally in living organisms. It
occurs at 66% relative humidity and in presence of lithium ions.
D-DNA - The D-form of DNA is found rarely as extreme vanants. It has 8 base pairs per turn of
helix. Therefore, it shows eight-fold symmetry. The base pairs are displaced backwardly with
respect to the axis of DNA helix.
E-DNA - E-DNA is found very rarely and the helix of E-DNA is long, and the bases are
perpendicular to the helical axis. It has 7.5 base pairs per helical turn. Environmental DNA or
E-DNA is DNA that is collected from a variety of environmental samples such as soil, seawater,
8
snow or air. E-DNA has been used as a tool to detect endangered wildlife that were otherwise
unseen.
[ऐसा पाया गया है क Watson और Crick structure क
े अलावा DNA क
ु छ अ य three dimensional forms
म भी हो सकता है I हालां क, इन variations से Watson & Crick वारा प रभा षत DNA क
े मुख गुण जैसे -
strand complementarity, antiparallel strands, और A=T तथा G≡C base pairing, पर कोई भाव नह ं
पड़ता क तु नकटवत base pairs क
े बीच angle of rotation म क
ु छ समायोजन हो सकता है I DNA
molecule क
े ये व भ न forms ह : A-DNA, B-DNA, Z-DNA, C-DNA, D-DNA, E-DNA
A-DNA - इसम 11 base pairs त helical turn होते ह I यह एक right handed double helix होता है क तु
थोड़ा अ धक चौड़ा होता है और bases helix क axis क
े ल बवत (perpendicular) होने क
े थान पर axis क
े
थोड़ा तरछे होते ह I इसम major groove थोड़ा गहर होती है क तु minor groove थोड़ा उथल (shallower)
होती है I यह form 75% relative humidity म पाया जाता है और इसक
े लए sodium, potassium या
caesium ions क उपि थ त आव यक होती है I
B-DNA - यह famous Watson & Crick structure है और living cells म पाया जाने वाला सबसे अ धक
सामा य संरचना (conformation) है I यह भी एक double stranded right handed helix होता है और
physiological conditions म सवा धक ि थर structure है I इसम 10.5 base pairs त helical turn होते ह I
इस प क
े लए अ धक humidity (92%) और low ionic strength क
े solutions क आव यकता होती है I
Z-DNA - यह एक left-handed double helix होता है िजसक
े sugar-phosphate backbone का pattern
zig-zag होता है I इसक संरचना right handed double helix वाले A-DNA और B-DNA से बहुत भ न होती
है I इसम 12 base pairs त helical turn होते ह और major groove मुि कल से ह तीत होता है तथा minor
groove संकरा (narrow) और गहरा होता है I Z-DNA बहुत से living organisms जैसे - mammals,
protozoans और कई plant species म पाया गया है I
C-DNA - यह भी DNA का एक double helical conformation है ले कन इसम 9.33 base pairs त helical
turn होते ह I DNA का यह प अ धक ि थर नह ं होता है और living organisms म वाभा वक प से नह ं पाया
जाता है I इसक
े लए 66% relative humidity और lithium ions क उपि थ त आव यक होती है I
D-DNA - DNA का यह प कभी कभार ह (अ य धक भ न प (extreme variant) क तरह पाया जाता है I
इसम 8 base pairs त helix turn होते ह, इस लए eight-fold symmetry दशाता है I इसम base pairs
DNA क axis क
े स दभ म थोड़ा पीछे क ओर खसक
े होते ह I
E-DNA - E-DNA भी बहुत मुि कल से मलता है I इसक helix ल बी होती है और bases helical axis क
े
ल बवत (perpendicular) होते ह I इसम त helical turn 7.5 base pairs होते ह I पयावरणीय डीएनए या
E-DNA वह DNA है जो व भ न कार क
े पयावरणीय नमून जैसे क म ट , समु जल, बफ या हवा से एक
कया जाता है I E-DNA का उपयोग लु त ाय व यजीव का पता लगाने क
े लए एक उपकरण क
े प म कया
गया है I
9
Differences between Z-DNA and B-DNA :
(a) The Z-DNA has a left handed helix, while the B-DNA has a right handed helix.
[Z-DNA म left handed helix होता है जब क B-DNA म यह right handed helix होता है I]
(b) The Z-DNA contains a zig-zag sugar-phosphate backbone as compared to regular backbone
of the B-DNA.
[Z-DNA म sugar-phosphate backbone zig-zag होती है जब क B-DNA म यह एक regular backbone
होती है I]
(c) The repeating unit in Z-DNA is a dinucleotide whereas in B-DNA the repeating unit is a
mononucleotide.
[Z-DNA क repeated units dinucleotides होती ह जब क B-DNA म ये mononucleotides क होती ह I]
(d) In the Z-DNA contains 12 base pairs per helical turn while in B-DNA consists of 10.5 base
pairs per helical turn.
[Z-DNA म 12 base pairs त helical turn होते ह जब क B-DNA म 10.5 base pairs त helical turn
होते ह I]
(e) Due to the presence of a higher number (12) of base pairs in one turn of Z-DNA, the angle of
twist per repeating unit i.e. dinucleotide is 60° as compared to 36° of B-DNA molecule.
[Z-DNA म त helical turn म अ धक base pairs (12) होने क
े कारण येक repeating dinucleotide
units पर angle of twist 60° होता है जब क B-DNA म यह angle 36° होता है I]
Q. 6 (A). Write a short note on DNA denaturation.[DNA क
े denaturation पर एक सं त ट पणी
लख I]
Ans. The process to break the double-stranded DNA into single-stranded by either physical
(heating) or chemical agents (high pH/ formaldehyde, urea) is known as DNA denaturation. In
the denaturation process, the hydrogen bonds between two strands are broken giving rise to
two single strands. The DNA denaturation process is reversible under controlled conditions of
pH and ionic strength. If the temperature is slowly decreased in the solution where the DNA had
been denatured, the DNA chains will spontaneously reanneal and the original double helix
structure is restored (renaturation). The temperature at which DNA is half denatured is called
10
critical temperature or melting temperature (Tm). These processes of DNA denaturation and
renaturation are utilized in polymerase chain reaction (PCR) technique.
**{Annealing is the process of joining of single-stranded DNA or RNA by hydrogen bonds to
form a double-stranded polynucleotide}
Apart from the structural changes in DNA the denaturation results in some changes in the
properties of DNA also:
● Increase in absorption of UV light - The absorbance (OD) of light at 260 nm increases
upon denaturation of DNA. It is due to unstacking of base pairs and this phenomenon is
called hyperchromicity or hyperchromatic effect.
● Decrease in viscosity - Denaturation causes a marked decrease in viscosity of DNA
solution.
These changes in the properties of DNA help in monitoring the progress of denaturation
process.
[ कसी physical (heating) तर क
े या chemical agents (high pH/ formaldehyde, urea) वारा
double-stranded DNA को single-stranded DNA म बदलने क या को DNA denaturation कहते ह I
इस या म DNA क
े दोन strands क
े बीच बने hydrogen bonds टूट जाते ह और इस कार दो single
strands ा त होते ह I pH और ionic strength क नयं त प रि थ तय म DNA denaturation एक
reversible या है I य द denatured DNA क
े solution का तापमान धीरे - धीरे घटाया जाए तो अलग हुई
DNA chains एनील (anneal) होकर पुनः double helix structure बना देती है और इस या को
renaturation कहते ह I िजस तापमान पर DNA आधा denature होता है उसे critical temperature या
melting temperature (Tm) कहते ह I DNA denaturation और renaturation क ये याओं का उपयोग
polymerase chain reaction (PCR) तकनीक म कया जाता है I
**{एनी लंग डबल- डेड पॉल यूि लयोटाइड बनाने क
े लए हाइ ोजन बांड वारा एकल- डेड डीएनए या
आरएनए को जोड़ने क या है। यह क
े वल पूरक अनु म क
े बीच होता है।}
DNA म structural changes क
े अ त र त denaturation क
े कारण DNA क properties म भी क
ु छ प रवतन
होते ह :
● UV light क
े absorption म वृ ध - DNA का denaturation होने पर 260 nm पर UV light का
absorbance (OD) म वृ ध होती है I ऐसा DNA क
े base pairs क
े अलग - अलग (unstacking) होने
क
े कारण होता है और इस प रघटना (phenomenon) को hyperchromicity या hyperchromatic
effect कहते ह I
● Viscosity म कमी - Denaturation क
े कारण DNA solution क viscosity कम हो जाती है I
DNA क properties म इन प रवतन का उपयोग denaturation process क ग त क नगरानी करने म
कया जाता है I]
11
Q.6 (B). What is hyperchromicity with respect to DNA?[DNA क
े स दभ म Hyperchromicity
या होती है?]
Ans. The absorption of light at 260 nm (UV light) is increased when the double stranded DNA is
denatured, either by heat or by addition of denaturant or by increasing the pH level, into two
single strands. This increase in absorbance of light is called hyperchromicity. In double stranded
DNA the two strands of DNA are bound together mainly by the stacking interactions, hydrogen
bonds and hydrophobic effect between the complementary bases. The denaturation results in
disruption of these interaction forces holding the double helical structure resulting in reduction of
base-base interaction and hence the absorbance of UV light at 260 nm by the resulting single
stranded DNA will be 37% higher than that for double stranded DNA at the same concentration.
Hyperchromicity can be used to track the condition of DNA e.g. on temperature changes i.e.
progress of denaturation of the DNA.
[जब double stranded DNA का heat या chemical agent वारा दो single strands म denaturation होता
है तो 260 nm पर light का absorption बढ़ जाता है I DNA वारा Light क
े absorbance म इस वृ ध को
hyperchromicity कहते ह I एक double stranded DNA क
े दोन strands उनक
े complementary bases क
े
बीच stacking interactions, hydrogen bonds और hydrophobic effect वारा बंधे रहते ह और
denaturation क
े कारण इन interactions क
े वघटन से bases क
े बीच interaction भी कम हो जाता है और
इस लए प रणाम व प बने single stranded DNA का 260 nm पर light absorbance double stranded
DNA क
े मुकाबले लगभग 37% अ धक होता है I इस hyperchromicity का उपयोग DNA क
े denaturation
process क ग त क नगरानी करने म कया जाता है I]
Q. 7. Write a short note on Transcription.[Transcription पर एक सं त ट पणी लख I]
Ans. Transcription is the process in which a segment of DNA sequence is copied (transcribed)
to make an RNA molecule. The RNA molecules so formed that can encode proteins are termed
as messenger RNA (mRNA). The mRNA, in turn, serves as a template for the protein's
synthesis through translation. Other segments of DNA may be transcribed into small non-coding
RNAs such as tRNA and rRNA.
The general steps of transcription :
● RNA polymerase, together with one or more general transcription factors, binds to
promoter sequence of DNA (transcription start sites) at the 5' end of DNA sequence.
● RNA polymerase generates a transcription bubble, which separates the two strands of
the DNA helix by breaking the hydrogen bonds between complementary DNA
nucleotides.
● RNA polymerase adds RNA nucleotides (ATP, GTP, CTP and UTP) complementary to
the nucleotides of one DNA strand.
● RNA sugar-phosphate backbone forms with assistance from RNA polymerase to form an
RNA strand.
● RNA polymerase encounters a terminator sequence and the transcription stops. RNA
polymerase then releases the DNA template.
● The transcribed RNA is processed further to convert it into mature RNA. RNA processing
includes capping, polyadenylation and splicing.
[Transcription वो या है िजसम DNA क
े एक भाग क
े sequence को copy करक
े एक RNA molecule
बनता है I इस कार बने RNA molecules जो protein synthesis क
े code रखते ह उ ह messenger RNA
(mRNA) कहते ह I ये mRNA फर व श ट protein क synthesis क
े लए एक सांचे (template) क तरह काम
12
करते ह और translation या वारा protein synthesis म भाग लेते ह I DNA क
े अ य भाग भी transcribe
होकर छोटे non-coding RNAs जैसे - tRNA और rRNA बनाते ह I
The general steps of transcription :
● RNA polymerase, एक या अ धक transcription factors क
े साथ, DNA क
े promoter sequence
(transcription start site) से bind करता है जो DNA sequence क
े 5' end पर होता है I
● RNA polymerase एक transcription bubble बनाता है जो DNA helix क
े दोन strands को पृथक
करता है और यह complementary DNA nucleotides क
े बीच क
े hydrogen bonds क
े टूटने क
े
कारण होता है I
● अब RNA polymerase एक DNA strand क
े nucleotides से complementary RNA nucleotides
(ATP, GTP, CTP and UTP) को जोड़ता है I
● RNA polymerase क सहायता से ह RNA sugar-phosphate backbone बन जाती है I
● अब RNA polymerase का एक terminator sequence से सामना होता है और transcription या
क जाती है I RNA polymerase तब DNA template को छोड़ देता है I
● यह transcribed RNA capping, polyadenylation और splicing जैसी याओं वारा mature
RNA म बदल जाता है I
Q. 8. Fill in the blanks -
(i) DNA molecule म दो वतं (independent) _________ strands होते ह I
(ii) DNA molecule क backbone phosphodiester bonds वारा जुड़े हुए _______ sugar units क एक
ृंखला से बनी होती है I
(iii) DNA म पाये जाने वाले purines ह ______ और ______
(iv) DNA म पाये जाने वाले pyrimidines ह ______ और ______
(v) DNA क
े एक strand क
े bases दूसरे strand क
े bases से ______ bonds वारा जुड़े होते ह I
(vi) DNA म base pairing हमेशा adenine (A) एवं thymine (T) क
े बीच ___ hydrogen bonds वारा और
cytosine(C) एवं guanine (G) क
े बीच ____ hydrogen bonds वारा होती है I
(vii) parent DNA क
े complementary DNA बनने क इस या को DNA ______ कहते ह I
(viii) RNA molecule म उपि थत pentose sugar का नाम है ________.
(ix) RNA म उपि थत चार nitrogenous bases है _____, _______, ______ और _____
(x) RNA म DNA म उपि थत Thymine क
े थान पर ________ होता है I
(xi) RNA _________ क
े single stranded polymers होते ह I
(xii) तीन कार क
े RNA ह - _______ RNA , _____ RNA और ______RNA
(xiii) mRNA म जो DNA क
े complementary bases क
े triplets होते ह उ ह _______ कहते ह I
(xiv) tRNA म उपि थत complementary bases क
े triplet को _______ कहते ह I
(xv) ______ म left handed double helix होता है I
(xvi) The process in which a segment of DNA sequence is copied to make an RNA molecule is
called ___________.
(xvii) inosine और pseudouridine tRNA म पाए जाने वाले ________ bases ह I
13
(xviii) कसी physical (heating) तर क
े या chemical agents (high pH/ formaldehyde, urea) वारा
double-stranded DNA को single-stranded DNA म बदलने क या को DNA _______ कहते ह I
(xix) िजस तापमान पर DNA आधा denature होता है उसे _______ temperature कहते ह I
(xx) DNA replication म इ तेमाल होने वाला enzyme ________________ है और transcription म
इ तेमाल होने वाला enzyme ___________ है I
Ans. (i) polydeoxyribonucleotide, (ii) deoxyribose, (iii) adenine, guanine, (iv) thymine, cytosine,
(v) hydrogen, (vi) दो, तीन, (vii) replication, (viii) ribose, (ix) adenine, guanine, cytosine uracil, (x)
uracil, (xi) ribonucleotides, (xii) ribosomal, messenger transfer, (xiii) codon, (xiv) anticodon, (xv)
Z-DNA, (xvi) transcription, (xvii) unusual, (xviii) denaturation, (xix) melting, (xx) DNA
polymerase, RNA polymerase
REFERENCES:
1. IGNOU, CHE - 9 Biochemistry, Block 1
2. Lehninger Principles of biochemistry, seventh edition ; David L. Nelson & Michael M. Cox.
Disclaimer : The pictures given in the text have been downloaded from Google images and I am
thankful to the persons who have uploaded these pictures.
Dr. P. K. Nigam (Retired Biochemist)

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Some Q & A Nucleic acids | Biochemistry IGNOU CHE-09 (in English /Hinglish)

  • 1. 1 4. SOME IMPORTANT QUESTION - ANSWER FOR IGNOU BIOCHEMISTRY [CHE 9] NUCLEIC ACIDS Q. 1. What are the principal constituents of nucleic acids? Mention the differences between a nucleoside and a nucleotide. Draw the structure of ATP.[Nucleic acids क े मु य अवयव या होते ह? Nucleoside और nucleotide म अंतर बताय I ATP क संरचना बनाएं ] Ans. Nucleic acids are polymerized chains of nucleotides (polynucleotides) joined by phosphodiester bonds. The two types of nucleic acids are deoxyribonucleic acid (DNA) and ribonucleic acids (RNA). Deoxyribonucleic acid (DNA) - is an universal genetic material and a fundamental unit of heredity. DNA consists of two independent polydeoxyribonucleotide strands in the form of a right handed double helix inside the cell. The backbone of DNA is made of a chain of deoxyribose sugar units joined together by phosphodiester bonds (between 3' end of one sugar and the 5' end of the next sugar). In addition, four types of nitrogeous bases [two purines - adenine (A) & guanine (G) and two pyrimidines - cytosine (C ) & thymine (T)] are also attached to sugar units and projected on inside of the backbone. The amount of A is equal to T, whereas the amount of C is equal to G (A=T; C=G). The bases of one strand are hydrogen bonded to the bases of the other strand of the double helix. The base pairing is always between adenine and thymine by two hydrogen bonds and between cytosine and guanine by three hydrogen bonds. In addition to these hydrogen bonds, there are some interactions like hydrophobic or stacking interactions which provide further stability to the double helix. Ribonucleic acids (RNA) - are single stranded polymers of ribonucleotides in which the pentose sugar is ribose instead of deoxyribose which is present in DNA. The ribose sugar units are joined together by phosphodiester bonds to form a strand to which four types of nitrogenous bases (adenine, guanine, cytosine and uracil) are attached. In RNA uracil nitrogenous base is present instead of thymine which is present in DNA. RNAs are generally single stranded except a few viral RNA molecules which are double stranded. The RNAs are the "middle man" of information transfer and there are three main types of RNAs based on the different roles played by them in the biosynthesis of proteins and polypeptides. These are ribosomal RNA (rRNA), messenger RNA (mRNA) and transfer RNA (tRNA). [Nucleic acids phodphodiester bonds वारा जुड़े nucleotides क polymerized chains होते ह अथात nucleotides क े polymer होते ह I दो कार क े nucleic acids ह - deoxyribonucleic acid (DNA) और ribonucleic acids (RNA) I Deoxyribonucleic acid (DNA) एक universal genetic material और आनुवं शकता (heredity) क मौ लक इकाई (fundamental unit) है I DNA molecule म दो वतं (independent) polydeoxyribonucleotide strands होते ह जो को शका क े अंदर एक right handed double helix क े प म होते ह I DNA molecule क backbone phosphodiester bonds वारा जुड़े हुए deoxyribose sugar units क एक ृंखला से बनी होती है और इसक े अ त र त इसम nitrogenous bases [two purines - adenine (A) & guanine (G) and two pyrimidines - cytosine (C ) & thymine (T)] भी जुड़े होते ह जो इस backbone से अंदर क ओर projected होते ह I A क मा ा T क े बराबर और C क मा ा G क े बराबर होती है (A=T; C=G) I एक strand क े bases दूसरे strand क े bases से hydrogen bonds वारा जुड़े होते ह और यह base pairing हमेशा adenine (A) एवं thymine (T) क े बीच दो hydrogen bonds वारा और cytosine (C) एवं guanine (G) क े बीच तीन
  • 2. 2 hydrogen bonds वारा होती है I इसक े अ त र त क ु छ अ य interactions जैसे hydrophobic और stacking interactions भी होते ह जो double helix को ि थरता दान करते ह I] Ribonucleic acid (RNA) - ये nucleotides क े single stranded polymer होते ह और इनम शुगर units ribose होती ह (DNA म deoxyribose sugar units होती ह) I DNA molecule क तरह ह RNA क backbone phosphodiester bonds वारा जुड़े हुए ribose sugar units क एक ृंखला से बनी होती है और इसक े अ त र त इसम nitrogenous bases [दो purines - adenine (A) एवं guanine (G) and दो pyrimidines - cytosine (C) एवं uracil (U)] भी जुड़े होते ह I RNAs सामा य तौर पर single stranded होते ह क तु क ु छ virus म ये double stranded होते ह I RNAs information transfer म बचौ लए (middle man) क भू मका नभाते ह I proteins और polypeptides क biosynthesis म अपनी -अपनी भू मका क े आधार पर RNA तीन कार क े होते ह - ribosomal RNA (rRNA), messenger RNA (mRNA) and transfer RNA (tRNA)] Difference between nucleoside and nucleotide: 1. A nucleoside consists of a sugar and a nitrogeous base without the phosphate group whereas a nucleotide consists of a phosphate group, a sugar and a nitrogenous base. [एक nucleoside एक sugar molecule और एक nitrogenous base से मलकर बना होता है और इसम कोई phosphate group नह ं होता है जब क nucleotide म sugar, nitrogenous base और एक phosphate group होता है] 2. The bond formed in a nucleoside is between C1 of the pentose sugar and either N1 of pyrimidine base or N9 of the purine base whereas in nucleotide a phosphate group is attached to C5 of the pentose sugar of the nucleoside by a phosphodiester linkage. [nucleoside म pentose sugar क े C1 और pyrimidine base क े N1 या purine base क े N9 क े बीच एक bond बना होता है जब क nucleotide म nucleoside क े pentose sugar क े C5 से एक phosphate group भी एक phodphodiester bond वारा जुड़ा होता है] Nucleosides Nucleotide
  • 3. 3 Structure of ATP Q. 2. Describe the salient features of Watson and Crick model of DNA. What are the important functions of DNA?[DNA क े Watson और Crick model क मु य वशेषताएं लख I DNA क े मु य काय लख ] Ans. Salient features of Watson and Crick model od DNA : ● DNA molecule has two independent polydeoxyribonucleotide strands which are in the form of a right handed double helix inside the cell. [DNA molecule म दो वतं (independent) polydeoxyribonucleotide strands होते ह जो को शका क े अंदर एक right handed double helix क े प म होते ह I] ● The two strands in DNA double helix are antiparallel, which means that the 5' end of one strand is paired with the 3' end of its complementary strand (and vice versa). [DNA क े double helix क े दोन strands antiparallel होते ह, अथात एक strand का 5' छोर दूसरे complementary strand क े 3' छोर क े साथ जोड़ी बनाता है ] ● The DNA backbone consists of a chain of deoxyribose sugar units joined by 3', 5'-phosphodiester bonds (between 3' end of one sugar and the 5' end of the next sugar) and these are projected outside of the backbone. [DNA molecule क backbone 3', 5'-phosphodiester bonds वारा जुड़े हुए deoxyribose sugar units क एक ृंखला से बनी होती है (between 3' end of one sugar and the 5' end of the next sugar) जो इस backbone क े बाहर क तरफ projected रहते ह ] ● In addition, two purine (adenine, guanine) and two pyrimidine (thymine, cytosine) bases are also attached to this backbone and projected inside of the backbone. [इसक े अ त र त इसम nitrogenous bases (two purines - adenine & guanine and two pyrimidines - cytosine & thymine) भी जुड़े होते ह जो इस backbone से अंदर क ओर projected होते ह] ● The bases of one strand are attached to the bases of the other strand by hydrogen bonds and this base pairing is always by two hydrogen bonds between adenine (A) & thymine (T) and by three hydrogen bonds between cytosine(C) & guanine (G). [एक strand क े bases दूसरे strand क े bases से hydrogen bonds वारा जुड़े होते ह और यह base pairing हमेशा adenine (A) एवं thymine (T) क े बीच दो hydrogen bonds वारा और cytosine(C) एवं guanine (G) क े बीच तीन hydrogen bonds वारा होती है ] ● A small groove, called the minor groove (1.2 nm), and a deep groove, known as the major groove (2.2 nm), run across the DNA helix which are created by the offset pairing of the two strands. [DNA क े दोन strands क ऑफसेट जोड़ी क े कारण पूरे DNA helix म small groove (1.2 nm) और deep groove (2.2 nm) होते ह ]
  • 4. 4 Functions of DNA - ● DNA is an universal genetic material and a fundamental unit of heredity. It is responsible for the inheritance and transmission of specific characteristics from one generation to the other. [DNA universal genetic material और आनुवं शकता (heredity) क मौ लक इकाई (fundamental unit) है I यह वंशानु म और एक पीढ़ से दूसर पीढ़ म व श ट वशेषताओं क े ह तांतरण क े लए िज़ मेदार होता है] ● DNA dictates how a human or animal develops and reproduces, and eventually dies. [DNA तय करता है क एक human या animal क ै से वक सत, जनन करता है और अंत म क ै से मृ यु को ा त होता है] ● DNA stores instructions for making other large molecules, called proteins. [DNA अ य बड़े molecules proteins क े नमाण क े लए वां छत नदश का सं ह करता है] ● DNA is also involved in [DNA इनम भी शा मल होता है] : (i) Equal distribution of DNA during the cell division [cell division क े समय DNA क े समान वतरण म ] (ii) Mutations i.e.The changes which occur in the DNA sequences [DNA sequence म होने वाले प रवतन म ] (iii) Transcription (RNA synthesis) [RNA क े सं लेषण (transcription) म ] (iv) Cellular Metabolism [cells क े अंदर होने वाल metabolism म ] (v) DNA fingerprinting [DNA fingerprinting म ] (vi) Gene Therapy [gene therapy म] Q. 3. What are the different types of RNA? Describe briefly.[ व भ न कार क े RNA या होते ह, सं ेप म वणन कर I] Ans. RNAs have been classified into three types viz. ribosomal RNA (rRNA), messenger RNA (mRNA) and transfer RNA (tRNA), depending on the basis of their different roles in biosynthesis of proteins and polypeptides. Ribosomal RNA - This constitutes about 66% of total RNA present in the ribosomes of a cell. It is synthesized in the nucleolus of the nucleus. These are non coding RNAs and are single stranded. The G:C and A:U ratios are unequal. In E. coli three forms of rRNA (5S, 16S, 23S) are found which differ in their base ratios and sequences. Likewise, in eukaryotic cells 5S, 7S, 18S and 28S forms of rRNAs are found. The formation of hydrogen bonds between base pairs results in the formation of the hairpin type turns in the rRNA molecules as found in tRNA. The precise role of rRNA is not clear but they are sites of polypeptide synthesis and they form complex with mRNA. The rRNA ensures the proper alignment of the mRNA, tRNA and the ribosomes. Messenger RNA - These are so called because they carry the 'message' from the coded DNA and execute/guide the synthesis of polypeptides. mRNAs are much shorter than rRNA and have
  • 5. 5 triplets of bases (codons) complementary to the DNA and each codon is the code for a particular amino acid. The sequence of these codons on the mRNA guides the synthesis of the particular polypeptide. The length of these mRNAs varies depending upon the lengths of polypeptides for which they are coded. mRNA is synthesized from DNA in the nucleus by a process called 'transcription' in which the genetic code (sequence of bases) is transcribed from one strand of the DNA molecule in the form of a single strand of mRNA with the help of an enzyme called RNA polymerase. After transcription, the mRNA passes into the cytoplasm and then to ribosomes where it serves as the template for the process of translation during protein synthesis. Transfer RNA - These are small molecules with 60-95 nucleotides. These are single stranded too but adopt a unique cloverleaf like structure due to extensive complementary base pairing of adenine:uracil and guanine:cytosine. Their function is to carry the individual amino acids to the site of protein synthesis and for this purpose they carry a complementary triplet of bases called the 'anticodon' which recognizes the codon of mRNA for a particular amino acid, thus acting as an interpreter in the process of translation in protein synthesis. The 3'- end ribose unit of tRNA has an OH group which can attach a particular amino acid by an ester bond forming aminoacyl-tRNA (charged form of tRNA). Each of the 20 amino acids found in proteins has at least one corresponding tRNA. The tRNA sometimes has unusual or minor bases like inosine, pseudouridine etc. The exact role of these unusual bases is not clearly known but these somehow protect the tRNA degradation by enzyme RNAase. Also these are involved in stabilization of the tertiary structure of tRNA. [RNA को protein और polypeptide synthesis म उनक भू मका क े आधार पर तीन कार म वग कृ त कया गया है - ribosomal RNA (rRNA), messenger RNA (mRNA) और transfer RNA (tRNA) I Ribosomal RNA - यह एक cell क े ribosomes म उपि थत क ु ल RNA का लगभग 66% होता है I इसका सं लेषण nucleus क े nucleolus म होता है I ये non-coding RNA होते ह और एक strand वाले होते ह I इनम G:C और A:U क े अनुपात असमान होते ह I E. coli म rRNA क े तीन प (5S, 16S, 23S) होते ह और इनम base ratios और sequences म भ नता होती है I इसी कार, eukaryotic cells म rRNA क े चार प (5S, 7S, 18S और 28S) पाए जाते ह I rRNA क े base pairs क े बीच hydrogen bonds बनने क े कारण एक hairpin type का प बन जाता है, जैसा tRNA म भी होता है I rRNA क ब क ु ल प ट भू मका क े बारे म प टता नह ं है क तु ये polypeptide synthesis क े काय थल होते ह और ये mRNA क े साथ complex बनाते ह I mRNA, tRNA और ribosomes क े बीच संरेखण (alignment) को rRNA सु नि चत करते ह I Messenger RNA - इ ह messenger RNA इस लए कहते ह य क ये DNA क े coded message को carry करने और polypeptide synthesis को न पा दत करने म भू मका नभाते ह I ये rRNA से काफ़ छोटे होते ह और इनम DNA क े complementary bases क े triplets (codons) होते ह और येक codon एक व श ट amino acid का code होता है I mRNA पर इन codons का जो sequence होता है वो एक नि चत polypeptide क synthesis का माग दशन करता है I इन mRNA क ल बाई उन polypeptides क ल बाई पर नभर करती है िजनक े लए ये mRNA coded होते ह I mRNA क synthesis nucleus म DNA से होती है और इस या को 'transcription' कहते ह िजसम DNA क े एक strand से genetic code (sequence of bases) को एक mRNA क े एक single strand क े प म अनुले खत (transcribe) कया जाता है, एक enzyme RNA polymerase क सहायता से I अनुलेखन (transcription) क े बाद यह mRNA cytoplasm म पहुँचता है और फर ribosomes पर पहुँचता है जहाँ यह protein synthesis क translation या म एक template क भू मका नभाता है I Transfer RNA - ये छोटे molecules होते ह िजनम 60-95 nucleotides होते ह I ये भी single stranded होते ह क तु adenine:uracil और guanine:cytosine क अ य धक base pairing क े कारण ये अनूठा cloverleaf
  • 6. 6 जैसी संरचना बना लेते ह I इनका काय होता है वैयि तक amino acids को protein synthesis क site पर ले जाना और इसक े लए इनम एक complementary bases का एक triplet होता है िजसे anticodon कहते ह I ये anticodon mRNA क े एक नि चत amino acid क े codon को पहचान कर protein synthesis क translation या म एक दुभा षया (interpreter) क भू मका नभाता है I tRNA क े 3'- end क े ribose unit क े OH group से एक नि चत amino acid एक ester bond वारा जुड़ कर aminoacyl-tRNA (charged form of tRNA) बनता है I proteins म पाए जाने वाले 20 amino acids म से येक amino acid का कम से कम एक अनु प (corresponding) tRNA होता है I कभी-कभी tRNA म unusual या minor bases जैसे - inosine, pseudouridine आ द भी पाए जाते ह I इन unusual bases क े काय क े बारे म प टता नह ं है क तु ये tRNA क े RNAase enzyme वारा कये जाने वाले अध:पतन (degradation) को कसी तरह रोकते ह I इसक े अ त र त ये tRNA क े tertiary structure को ि थरता दान करने म भी भू मका नभाते ह I Q. 4. What are unusual bases? Write the major differences between RNA and DNA.[असामा य bases या होते ह? RNA और DNA म मु य अंतर लख I] Ans. Unusual bases - In addition to primary nucleobases, DNA and RNA also contain other (non-primary) unusual bases that have been modified after the nucleic acid chain has been formed. These are formed by enzymatic modifications of the existing polynucleotides. In DNA, the most common modified bases are 5-methylcytosine (m5C) in animals and higher plants, N6 methyladenosine in bacterial DNA and 5-hydroxymethylcytosine in animals and bacteria infected with certain bacteriophages. The minor or unusual bases found in RNA, especially in tRNA are Inosine, 1-methylinosine, N2-dimethylguanosine, ribothymidine, dihydrouridine, pseudouridine, 7-methylguanosine and 4-thiouridine. The exact role of these unusual bases is not clearly known. These somehow protect the tRNA degradation by RNAase. Also these are involved in stabilization of the tertiary structure of tRNA. In DNA these unusual bases somehow play some role in regulating or protecting the genetic information. [DNA और RNA म सामा य (primary) nucleobases क े अ त र त क ु छ असामा य (unusual/non-primary) bases भी पाए जाते ह जो nucleic acid chain बनने क े बाद modify होकर बनते ह और ये पहले से मौजूद nucleotides क े enzymatic modifications वारा बनते ह I DNA म आमतौर पाए जाने वाले modified bases ह - 5-methylcytosine (m5C) animals और higher plants म , N6 methyladenosine bacterial DNA म और 5-hydroxymethylcytosine animals और bacteria म जो क ु छ bacteriophages से सं मत ह I RNA म आमतौर पर पाए जाने वाले unusual या असामा य bases, वशेषतः tRNA म, ह - Inosine, 1-methylinosine, N2-dimethylguanosine, ribothymidine, dihydrouridine, pseudouridine, 7-methylguanosine और 4-thiouridine I इन unusual bases क े नि चत काय क े बारे म प ट जानकार नह ं है क तु ये tRNA क े RNAase enzyme वारा कये जाने वाले अध:पतन (degradation) को कसी तरह रोकते ह और tRNA क े tertiary structure को ि थरता दान करने म भी भू मका नभाते ह I DNA म ये unusual bases DNA क genetic information क े नयं ण और संर ण म क ु छ भू मका नभाते ह ]
  • 7. 7 Difference between DNA and RNA - ● DNA contains deoxyribose sugar but RNA contains ribose sugar [DNA म शुगर deoxyribose होती है और RNA म ribose sugar होती है ] ● DNA is a double stranded molecule but RNA is a single stranded molecule. [DNA म दो strands होते ह जब क RNA म एक strand होता है ] ● DNA contains adenine, guanine, cytosine and thymine but RNA contains uracil in place of thymine. [DNA म adenine, guanine, cytosine और thymine bases होते ह जब क RNA म thymine क े थान पर uracil base होता है ] ● DNA is self replicating but RNA is synthesized by transcription of DNA. [DNA वयं से replicate होता है (self replicating) जब क RNA DNA क े transcription से बनता है ] ● DNA contains genetic information for the cell but RNA uses the information stored in DNA to make proteins. [DNA म genetic information न हत रहती है जब क RNA DNA क इस information को protein synthesis म उपयोग करता है ] Q. 5. What are the different forms of DNA? Mention the main differences between the B- DNA and Z-DNA.[DNA क े व भ न प या होते ह? B-DNA और Z-DNA म मु य अंतर या ह?] Ans. It has been found that DNA can occur in different three dimensional forms other than Watson and Crick DNA structure. However, these variations generally do not affect the key properties of DNA defined by Watson and Crick such as strand complementarity, antiparallel strands, and the requirement for A=T and G≡C base pairing but there may be slight adjustment in the angle of rotation between the adjacent base pairs. The different forms of DNA molecules are: A-DNA, B-DNA, Z-DNA, C-DNA, D-DNA, E-DNA A-DNA - has 11 base pairs per helical turn which is more than the average. The double helix is right handed but a little wider but bases lie tilted to the helix axis instead of perpendicular to the helix The major groove is deeper but minor groove is shallower. This form appears at 75% relative humidity and require the presence of sodium, potassium or caesium ions. B-DNA - It is the famous Watson & Crick structure and is the most common conformation in the living cells and is a right handed helix. It is the most stable structure under physiological conditions. It has 10.5 base pairs per helical turn. It requires high humidity (92%) and solutions of low ionic strength. Z-DNA - It is a left-handed double helix with a zig-zag pattern in the sugar-phosphate backbone. It has a very different structure when compared with A-DNA and B-DNA which are right-handed double helix structures. It has 12 base pairs per helical turn. The major groove is barely apparent and the minor groove is narrow and deep. The Z-DNA has been found in a large number of living organisms including mammals, protozoans and several plant species. C-DNA - It is also a double helical conformations of DNA but has 9.33 base pairs per helical turn. The C-form DNA is not very stable and does not occur naturally in living organisms. It occurs at 66% relative humidity and in presence of lithium ions. D-DNA - The D-form of DNA is found rarely as extreme vanants. It has 8 base pairs per turn of helix. Therefore, it shows eight-fold symmetry. The base pairs are displaced backwardly with respect to the axis of DNA helix. E-DNA - E-DNA is found very rarely and the helix of E-DNA is long, and the bases are perpendicular to the helical axis. It has 7.5 base pairs per helical turn. Environmental DNA or E-DNA is DNA that is collected from a variety of environmental samples such as soil, seawater,
  • 8. 8 snow or air. E-DNA has been used as a tool to detect endangered wildlife that were otherwise unseen. [ऐसा पाया गया है क Watson और Crick structure क े अलावा DNA क ु छ अ य three dimensional forms म भी हो सकता है I हालां क, इन variations से Watson & Crick वारा प रभा षत DNA क े मुख गुण जैसे - strand complementarity, antiparallel strands, और A=T तथा G≡C base pairing, पर कोई भाव नह ं पड़ता क तु नकटवत base pairs क े बीच angle of rotation म क ु छ समायोजन हो सकता है I DNA molecule क े ये व भ न forms ह : A-DNA, B-DNA, Z-DNA, C-DNA, D-DNA, E-DNA A-DNA - इसम 11 base pairs त helical turn होते ह I यह एक right handed double helix होता है क तु थोड़ा अ धक चौड़ा होता है और bases helix क axis क े ल बवत (perpendicular) होने क े थान पर axis क े थोड़ा तरछे होते ह I इसम major groove थोड़ा गहर होती है क तु minor groove थोड़ा उथल (shallower) होती है I यह form 75% relative humidity म पाया जाता है और इसक े लए sodium, potassium या caesium ions क उपि थ त आव यक होती है I B-DNA - यह famous Watson & Crick structure है और living cells म पाया जाने वाला सबसे अ धक सामा य संरचना (conformation) है I यह भी एक double stranded right handed helix होता है और physiological conditions म सवा धक ि थर structure है I इसम 10.5 base pairs त helical turn होते ह I इस प क े लए अ धक humidity (92%) और low ionic strength क े solutions क आव यकता होती है I Z-DNA - यह एक left-handed double helix होता है िजसक े sugar-phosphate backbone का pattern zig-zag होता है I इसक संरचना right handed double helix वाले A-DNA और B-DNA से बहुत भ न होती है I इसम 12 base pairs त helical turn होते ह और major groove मुि कल से ह तीत होता है तथा minor groove संकरा (narrow) और गहरा होता है I Z-DNA बहुत से living organisms जैसे - mammals, protozoans और कई plant species म पाया गया है I C-DNA - यह भी DNA का एक double helical conformation है ले कन इसम 9.33 base pairs त helical turn होते ह I DNA का यह प अ धक ि थर नह ं होता है और living organisms म वाभा वक प से नह ं पाया जाता है I इसक े लए 66% relative humidity और lithium ions क उपि थ त आव यक होती है I D-DNA - DNA का यह प कभी कभार ह (अ य धक भ न प (extreme variant) क तरह पाया जाता है I इसम 8 base pairs त helix turn होते ह, इस लए eight-fold symmetry दशाता है I इसम base pairs DNA क axis क े स दभ म थोड़ा पीछे क ओर खसक े होते ह I E-DNA - E-DNA भी बहुत मुि कल से मलता है I इसक helix ल बी होती है और bases helical axis क े ल बवत (perpendicular) होते ह I इसम त helical turn 7.5 base pairs होते ह I पयावरणीय डीएनए या E-DNA वह DNA है जो व भ न कार क े पयावरणीय नमून जैसे क म ट , समु जल, बफ या हवा से एक कया जाता है I E-DNA का उपयोग लु त ाय व यजीव का पता लगाने क े लए एक उपकरण क े प म कया गया है I
  • 9. 9 Differences between Z-DNA and B-DNA : (a) The Z-DNA has a left handed helix, while the B-DNA has a right handed helix. [Z-DNA म left handed helix होता है जब क B-DNA म यह right handed helix होता है I] (b) The Z-DNA contains a zig-zag sugar-phosphate backbone as compared to regular backbone of the B-DNA. [Z-DNA म sugar-phosphate backbone zig-zag होती है जब क B-DNA म यह एक regular backbone होती है I] (c) The repeating unit in Z-DNA is a dinucleotide whereas in B-DNA the repeating unit is a mononucleotide. [Z-DNA क repeated units dinucleotides होती ह जब क B-DNA म ये mononucleotides क होती ह I] (d) In the Z-DNA contains 12 base pairs per helical turn while in B-DNA consists of 10.5 base pairs per helical turn. [Z-DNA म 12 base pairs त helical turn होते ह जब क B-DNA म 10.5 base pairs त helical turn होते ह I] (e) Due to the presence of a higher number (12) of base pairs in one turn of Z-DNA, the angle of twist per repeating unit i.e. dinucleotide is 60° as compared to 36° of B-DNA molecule. [Z-DNA म त helical turn म अ धक base pairs (12) होने क े कारण येक repeating dinucleotide units पर angle of twist 60° होता है जब क B-DNA म यह angle 36° होता है I] Q. 6 (A). Write a short note on DNA denaturation.[DNA क े denaturation पर एक सं त ट पणी लख I] Ans. The process to break the double-stranded DNA into single-stranded by either physical (heating) or chemical agents (high pH/ formaldehyde, urea) is known as DNA denaturation. In the denaturation process, the hydrogen bonds between two strands are broken giving rise to two single strands. The DNA denaturation process is reversible under controlled conditions of pH and ionic strength. If the temperature is slowly decreased in the solution where the DNA had been denatured, the DNA chains will spontaneously reanneal and the original double helix structure is restored (renaturation). The temperature at which DNA is half denatured is called
  • 10. 10 critical temperature or melting temperature (Tm). These processes of DNA denaturation and renaturation are utilized in polymerase chain reaction (PCR) technique. **{Annealing is the process of joining of single-stranded DNA or RNA by hydrogen bonds to form a double-stranded polynucleotide} Apart from the structural changes in DNA the denaturation results in some changes in the properties of DNA also: ● Increase in absorption of UV light - The absorbance (OD) of light at 260 nm increases upon denaturation of DNA. It is due to unstacking of base pairs and this phenomenon is called hyperchromicity or hyperchromatic effect. ● Decrease in viscosity - Denaturation causes a marked decrease in viscosity of DNA solution. These changes in the properties of DNA help in monitoring the progress of denaturation process. [ कसी physical (heating) तर क े या chemical agents (high pH/ formaldehyde, urea) वारा double-stranded DNA को single-stranded DNA म बदलने क या को DNA denaturation कहते ह I इस या म DNA क े दोन strands क े बीच बने hydrogen bonds टूट जाते ह और इस कार दो single strands ा त होते ह I pH और ionic strength क नयं त प रि थ तय म DNA denaturation एक reversible या है I य द denatured DNA क े solution का तापमान धीरे - धीरे घटाया जाए तो अलग हुई DNA chains एनील (anneal) होकर पुनः double helix structure बना देती है और इस या को renaturation कहते ह I िजस तापमान पर DNA आधा denature होता है उसे critical temperature या melting temperature (Tm) कहते ह I DNA denaturation और renaturation क ये याओं का उपयोग polymerase chain reaction (PCR) तकनीक म कया जाता है I **{एनी लंग डबल- डेड पॉल यूि लयोटाइड बनाने क े लए हाइ ोजन बांड वारा एकल- डेड डीएनए या आरएनए को जोड़ने क या है। यह क े वल पूरक अनु म क े बीच होता है।} DNA म structural changes क े अ त र त denaturation क े कारण DNA क properties म भी क ु छ प रवतन होते ह : ● UV light क े absorption म वृ ध - DNA का denaturation होने पर 260 nm पर UV light का absorbance (OD) म वृ ध होती है I ऐसा DNA क े base pairs क े अलग - अलग (unstacking) होने क े कारण होता है और इस प रघटना (phenomenon) को hyperchromicity या hyperchromatic effect कहते ह I ● Viscosity म कमी - Denaturation क े कारण DNA solution क viscosity कम हो जाती है I DNA क properties म इन प रवतन का उपयोग denaturation process क ग त क नगरानी करने म कया जाता है I]
  • 11. 11 Q.6 (B). What is hyperchromicity with respect to DNA?[DNA क े स दभ म Hyperchromicity या होती है?] Ans. The absorption of light at 260 nm (UV light) is increased when the double stranded DNA is denatured, either by heat or by addition of denaturant or by increasing the pH level, into two single strands. This increase in absorbance of light is called hyperchromicity. In double stranded DNA the two strands of DNA are bound together mainly by the stacking interactions, hydrogen bonds and hydrophobic effect between the complementary bases. The denaturation results in disruption of these interaction forces holding the double helical structure resulting in reduction of base-base interaction and hence the absorbance of UV light at 260 nm by the resulting single stranded DNA will be 37% higher than that for double stranded DNA at the same concentration. Hyperchromicity can be used to track the condition of DNA e.g. on temperature changes i.e. progress of denaturation of the DNA. [जब double stranded DNA का heat या chemical agent वारा दो single strands म denaturation होता है तो 260 nm पर light का absorption बढ़ जाता है I DNA वारा Light क े absorbance म इस वृ ध को hyperchromicity कहते ह I एक double stranded DNA क े दोन strands उनक े complementary bases क े बीच stacking interactions, hydrogen bonds और hydrophobic effect वारा बंधे रहते ह और denaturation क े कारण इन interactions क े वघटन से bases क े बीच interaction भी कम हो जाता है और इस लए प रणाम व प बने single stranded DNA का 260 nm पर light absorbance double stranded DNA क े मुकाबले लगभग 37% अ धक होता है I इस hyperchromicity का उपयोग DNA क े denaturation process क ग त क नगरानी करने म कया जाता है I] Q. 7. Write a short note on Transcription.[Transcription पर एक सं त ट पणी लख I] Ans. Transcription is the process in which a segment of DNA sequence is copied (transcribed) to make an RNA molecule. The RNA molecules so formed that can encode proteins are termed as messenger RNA (mRNA). The mRNA, in turn, serves as a template for the protein's synthesis through translation. Other segments of DNA may be transcribed into small non-coding RNAs such as tRNA and rRNA. The general steps of transcription : ● RNA polymerase, together with one or more general transcription factors, binds to promoter sequence of DNA (transcription start sites) at the 5' end of DNA sequence. ● RNA polymerase generates a transcription bubble, which separates the two strands of the DNA helix by breaking the hydrogen bonds between complementary DNA nucleotides. ● RNA polymerase adds RNA nucleotides (ATP, GTP, CTP and UTP) complementary to the nucleotides of one DNA strand. ● RNA sugar-phosphate backbone forms with assistance from RNA polymerase to form an RNA strand. ● RNA polymerase encounters a terminator sequence and the transcription stops. RNA polymerase then releases the DNA template. ● The transcribed RNA is processed further to convert it into mature RNA. RNA processing includes capping, polyadenylation and splicing. [Transcription वो या है िजसम DNA क े एक भाग क े sequence को copy करक े एक RNA molecule बनता है I इस कार बने RNA molecules जो protein synthesis क े code रखते ह उ ह messenger RNA (mRNA) कहते ह I ये mRNA फर व श ट protein क synthesis क े लए एक सांचे (template) क तरह काम
  • 12. 12 करते ह और translation या वारा protein synthesis म भाग लेते ह I DNA क े अ य भाग भी transcribe होकर छोटे non-coding RNAs जैसे - tRNA और rRNA बनाते ह I The general steps of transcription : ● RNA polymerase, एक या अ धक transcription factors क े साथ, DNA क े promoter sequence (transcription start site) से bind करता है जो DNA sequence क े 5' end पर होता है I ● RNA polymerase एक transcription bubble बनाता है जो DNA helix क े दोन strands को पृथक करता है और यह complementary DNA nucleotides क े बीच क े hydrogen bonds क े टूटने क े कारण होता है I ● अब RNA polymerase एक DNA strand क े nucleotides से complementary RNA nucleotides (ATP, GTP, CTP and UTP) को जोड़ता है I ● RNA polymerase क सहायता से ह RNA sugar-phosphate backbone बन जाती है I ● अब RNA polymerase का एक terminator sequence से सामना होता है और transcription या क जाती है I RNA polymerase तब DNA template को छोड़ देता है I ● यह transcribed RNA capping, polyadenylation और splicing जैसी याओं वारा mature RNA म बदल जाता है I Q. 8. Fill in the blanks - (i) DNA molecule म दो वतं (independent) _________ strands होते ह I (ii) DNA molecule क backbone phosphodiester bonds वारा जुड़े हुए _______ sugar units क एक ृंखला से बनी होती है I (iii) DNA म पाये जाने वाले purines ह ______ और ______ (iv) DNA म पाये जाने वाले pyrimidines ह ______ और ______ (v) DNA क े एक strand क े bases दूसरे strand क े bases से ______ bonds वारा जुड़े होते ह I (vi) DNA म base pairing हमेशा adenine (A) एवं thymine (T) क े बीच ___ hydrogen bonds वारा और cytosine(C) एवं guanine (G) क े बीच ____ hydrogen bonds वारा होती है I (vii) parent DNA क े complementary DNA बनने क इस या को DNA ______ कहते ह I (viii) RNA molecule म उपि थत pentose sugar का नाम है ________. (ix) RNA म उपि थत चार nitrogenous bases है _____, _______, ______ और _____ (x) RNA म DNA म उपि थत Thymine क े थान पर ________ होता है I (xi) RNA _________ क े single stranded polymers होते ह I (xii) तीन कार क े RNA ह - _______ RNA , _____ RNA और ______RNA (xiii) mRNA म जो DNA क े complementary bases क े triplets होते ह उ ह _______ कहते ह I (xiv) tRNA म उपि थत complementary bases क े triplet को _______ कहते ह I (xv) ______ म left handed double helix होता है I (xvi) The process in which a segment of DNA sequence is copied to make an RNA molecule is called ___________. (xvii) inosine और pseudouridine tRNA म पाए जाने वाले ________ bases ह I
  • 13. 13 (xviii) कसी physical (heating) तर क े या chemical agents (high pH/ formaldehyde, urea) वारा double-stranded DNA को single-stranded DNA म बदलने क या को DNA _______ कहते ह I (xix) िजस तापमान पर DNA आधा denature होता है उसे _______ temperature कहते ह I (xx) DNA replication म इ तेमाल होने वाला enzyme ________________ है और transcription म इ तेमाल होने वाला enzyme ___________ है I Ans. (i) polydeoxyribonucleotide, (ii) deoxyribose, (iii) adenine, guanine, (iv) thymine, cytosine, (v) hydrogen, (vi) दो, तीन, (vii) replication, (viii) ribose, (ix) adenine, guanine, cytosine uracil, (x) uracil, (xi) ribonucleotides, (xii) ribosomal, messenger transfer, (xiii) codon, (xiv) anticodon, (xv) Z-DNA, (xvi) transcription, (xvii) unusual, (xviii) denaturation, (xix) melting, (xx) DNA polymerase, RNA polymerase REFERENCES: 1. IGNOU, CHE - 9 Biochemistry, Block 1 2. Lehninger Principles of biochemistry, seventh edition ; David L. Nelson & Michael M. Cox. Disclaimer : The pictures given in the text have been downloaded from Google images and I am thankful to the persons who have uploaded these pictures. Dr. P. K. Nigam (Retired Biochemist)