Gold Notes of Mol Bio by Garima Mam

MOLECULAR BASIS OF
INHERITANCE
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NUCLEIC ACID (DNA AND RNA)
Discovered by :- F. Meischer (1869)
Polymer of Nucleotides
Nucleotide :-Nitrogen Base + Sugar + Phosphate

Nitrogen base –two types :-
1. Pyrimidines (C, U, T)
2. Purines (A, G)

PYRIMIDINES

PURINES

PENTOSE SUGAR

PHOSPHATE
1. Acidic
2. Negative charge

STRUCTURE OF DNA
Wilkins and Frankline –X –ray crystallography
Watson and Crick –Double Helix Model
DNA
5 3
1. Double stand
’ ’
2. Antiparallel
3. Complimentary
4. H bonds (A =T, G=C)
5. Uniform diameter (20Å)
3 5 6. Distance between two base pair 3.4Å

’ 7. Helix length –34Å
’
8. Steps –base pair
Diameter = 20 Å
9. Backbone –sugar + phosphate

CHARGAFF’S RULE
Purine = Pyrimidine
A+G=C+T
A + G/ C + T = 1
Base pairs in some organisms

PACKAGING OF DNA HELIX
In eukaryotes packaging is
Carried out by histone protein :-
1. Basic protein
2. Lysine and arginine rich
A typical nucleosome contain 200 base pairs
Nucleosome

SEARCH FOR GENETIC MATERIAL
Transforming Principle :-Griffith
Worked on streptococcus pneumonia
1. S III Virulent
Two strain :-
2. R II nonvirulent
Heat killed S III + Live R II + Mice = Mice dead

(R II converted into S III)

SEARCH FOR GENETIC MATERIAL
Biochemical characterization of Transforming Principle
: by Avery, Macleod and McCarty

(proved that Transformation was due to DNA)
Final proved that DNA is genetic material by :-
Hershey and Chase (1952) :-
1. Worked with T2 bacteriophage

2. Use isotope of S(S35) and
P(P32)

PROPERTIES OF GENETIC MATERIAL
(1) The genetic material should be able to Replicate.
DNA (yes)
RNA (yes)
PROTEIN (no)
(2) It should chemically and structurally be stable
DNA more stable :-Presence of deoxyribose sugar

Presence of Thymine
(3) The genetic material should also be capable of undergoing slow mutations
(4) The genetic material should be able express as Mendelian characters

DNA REPLICATION
 In eukaryotes take place in:-S phase
 In prokaryotes take pace in :-Just Prior to fission Method
 :-Semi Conservative
 Idea :-Watson and Crick
 Proved by :-Meselson and Stahl
 Worked on :-E. coli bacteria
 Use isotopof Nitrogen (N14, N15)
 Proved at chromosome level :Taylor
 Worked on :-Viccia fava
 Use radioactive thymidine

MECHANISM OF DNA REPLICATION
Step 1 :-Unzipping (Unwinding)
 The process of unzipping starts at a certain specific point called origin of replication (ori).
 Enzyme (protein):
 Helicase:-Breaks hydrogen bonds
 SSB Protein :-Prevents the reformation of H bonds
 Topoisomerase :-Prevent supercoiling

Step 2 :-Formation of new chain :-always in 5’ 3’

Main enzyme :-
 DNA dependent DNA polymerase
 DNA polymerase I :-(a) Remove RNA primer (b) Gap filling DNA
 polymerase II :-Least role

 DNA polymerase III :-Main enzyme
 Primase :-(a) From RNA Primer
(b) Starting or replication
 DNA Ligase :-Join DNA fragments

Replication fork
lagging or
5 discontinuous strand
3
’
Leading ’
Or
Continuous strand
3
5
’
’
r-RNA
80% of the cell’s total RNA (most abundant RNA)
Most stable form of RNA
Function :-
1. Form structure of ribosomes
2. Some r-NA act as a catyulast / enzyme
(Ribozyme) e.g. 23 s r –RNA

t-RNA
 Soluble RNA
 Adapter (carrier) RNA
 Smallest RNA
 Have anticodon
Structure :-
2D :-Clover leaf like
3D :-Inverted L
Function :-
Transfer of amino –acids

TRANSCRIPTION (DNA  RNA)
5’ SENSE
OR
CODING STRAND
3’
RNA DNA
3’
5’
ANTTISENSE
5’
OR
TEMPLATE STRAND
3’
TRANSCRIPTION
 ENZYME :-
DNA dependent RNA polymerase

TRANSCRIPTION
 Eukaryotes :-three types of RNA polymerase
RNA polymerase I 28s RNA, 18s rRNA, 5.8S rRNA
RNA polymerase II hn –RNA, m-RNA

t-RNA,5s-rRNA, SnRNA
RNApolymerase III

TRANSCRIPTION UNIT

TRANSCRIPTION

CHARGFF’S RULE
FOUND ONLY IN EUKARYOTES
5’ EXONINTRONEXON3’
Hn RNA
1. Unstable
2. Contain intron
7 Methyl guanosine 5’ EXON INTRON EXON 3’

AAAAAAAAA-------
Tri phosphate
POLY ‘A’ TAIL (TAILING)
CAPPING
SPLICING BY SPLICEOSOME
COMPLEX
7 Methyl guanosine 5’ EXON EXON 3’

AAAAAAAAA-------
Tri phosphate
m-RNA

GENETIC CODE
1. Triplet in nature
2. Universal
3. Non overlapping
4. Commaless
5. Non ambiguous :-1 codon code 1 amino acid

6. Degeneracy of genetic code :-
1. amino acid code by more than 1 codon

GENETIC CODE
 Start / Initiation codon:- AUG
 Stop / nonsense codon :- UAA
UAG
UGA

TRANSLATION (PROTEIN SYNTHESIS)
1. Activation of amino acid :-
Enzyme Amino acyl AMP
Amino acid + ATP
Enzyme complex
2. Charging of t-RNA
Amino acyl AMP

+ t-RNA Amino acyl t-RNA
Enzyme complex complex

3. Transition :-3 steps
(A) Initiation :-

(B) Elongation :-

TRANSLATION (PROTEINS SYNTHESIS)
(C) Termination :-

GENE REGULATION (LAC OPERON)
 Lac : -lactose
 Beta –galactosidase :-break lactose into glucose and galactose
 Permease :-enter lactose in to the cell
 Catabolic pathway
 Inducible operon (off on)

 Inducer :-lactose
 Rea inducer :-allolactose
 Show both Negative and Positive regulation

 Regulation by repressor protein :-Negative

GENE REGULATION (LAC OPERON)
Binding site for
Binding site for
Repressor protein
RNA Polymerase
Operator
Promoter
P i P O z y a
𝜷 -Galactosidase 𝑷𝒆𝒓𝒎𝒆𝒂𝒔𝒆 𝑻𝒓𝒂𝒏𝒔𝒂𝒄𝒆𝒕𝒚𝒍𝒂𝒔𝒆
Repressor
protein

HUMAN GENOME PROJECT (HGP)
 Started in 1990
 Competed in 2003
 Chromosome 1st MAY 2006

 Studied 24 chromosomes
 Mega project
 Vector :-BAC and YAC
 Instrument :-Automated DNA sequence based on SANER principle
 Method :-
1.EST (Expressed Sequence Tag) :-Identifying all the genes that expressed as RNA
2.Sequence Annotation :-Sequencing the whole set of exome (coding and non-coding sequence)

HUMAN GENOME PROJECT (HGP)
Feature :-
1. Human genome contain :-3164.7 bases
2. Average gene contain :-3000 bases
3. Largest gene :-Dystrophin 2.4 million bases

4. Estimated genes :-30000
5. Function unknown :-Over 50% genes 6.
Coding part :_ Less than 2%

7. Most gene :-Chromosome 1 (2968)
8. Fewest genes :-Chromosome Y (231)
9. SNPs :-1.4 million locations

10. Repetitive sequence :-Large portion of human genome
NCERT FIG

DNA FINGERPRINTING
 DNA of human is almost (99.9%) same but very small amount (0.1%)
that differs from person to person

This differences are mainly at –Repetive DNA
• Do not code for any protein
• Make up large portion of human genome
• Show high degree of polymorphism(variation) so basis of DNA fingerprinting

• Mainly present in minor peaks (satiate DNA)
• Satellite DNA :-e.g. VNTR
• VNTR :-Variable Number of Tandem Repeat
-Type of mini satellite
• -Size –0.1kb to 20 kb

DNA FINGERPRINTING
Isolation of DNA cut by Restriction Separation of DNA
DNA Endonuclease Fragments by gel
electrophoresis
Denaturation by
Alkaline medium
Southern Botting
Autoradiography Hybridization using labelled

(Use X-ray film) VNTR Probe


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MOLECULAR BASIS OF

Telegram:https://t.me/garimagoel007_g2 GOLD NOTES BY GARIMA GOEL

Nucleotide :-Nitrogen Base + Sugar + Phosphate

Telegram:https://t.me/garimagoel007_g2 GOLD NOTES BY GARIMA GOEL

Telegram:https://t.me/garimagoel007_g2 GOLD NOTES BY GARIMA GOEL

Telegram:https://t.me/garimagoel007_g2 GOLD NOTES BY GARIMA GOEL

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3 5 6. Distance between two base pair 3.4Å

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2. Lysine and arginine rich

A typical nucleosome contain 200 base pairs

Telegram:https://t.me/garimagoel007_g2 GOLD NOTES BY GARIMA GOEL

Worked on streptococcus pneumonia

Heat killed S III + Live R II + Mice = Mice dead

Telegram:https://t.me/garimagoel007_g2 GOLD NOTES BY GARIMA GOEL

: by Avery, Macleod and McCarty

Final proved that DNA is genetic material by :-

Hershey and Chase (1952) :-

1. Worked with T2 bacteriophage

Telegram:https://t.me/garimagoel007_g2 GOLD NOTES BY GARIMA GOEL

DNA more stable :-Presence of deoxyribose sugar

(4) The genetic material should be able express as Mendelian characters

Telegram:https://t.me/garimagoel007_g2 GOLD NOTES BY GARIMA GOEL

 In prokaryotes take pace in :-Just Prior to ﬁssion Method

 Idea :-Watson and Crick

 Proved by :-Meselson and Stahl

 Worked on :-E. coli bacteria

 Use isotopof Nitrogen (N14, N15)

 Proved at chromosome level :Taylor

 Worked on :-Viccia fava

 Use radioactive thymidine

 Helicase:-Breaks hydrogen bonds

 SSB Protein :-Prevents the reformation of H bonds

 Topoisomerase :-Prevent supercoiling

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 polymerase II :-Least role

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Most stable form of RNA

(Ribozyme) e.g. 23 s r –RNA

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 Adapter (carrier) RNA

2D :-Clover leaf like

Transfer of amino –acids

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DNA dependent RNA polymerase

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 Eukaryotes :-three types of RNA polymerase

RNA polymerase I 28s RNA, 18s rRNA, 5.8S rRNA

RNA polymerase II hn –RNA, m-RNA

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7 Methyl guanosine 5’ EXON INTRON EXON 3’

7 Methyl guanosine 5’ EXON EXON 3’

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5. Non ambiguous :-1 codon code 1 amino acid

1. amino acid code by more than 1 codon

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 Stop / nonsense codon :- UAA

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Amino acyl AMP