AVERY’S EXPERIMENT

Removal of all protein from transforming material did not destroy its
ability to transform R strain cells

DNA-digesting enzyme destroyes all transforming ability

The transforming material is DNA

STEPS:-

 Cell free extract of SIII strain bacterium was subjected to Dnase,

Rnase and protease
 Each treated extract was mixed with RII and mixture injected to
mouse to see transformation
 Incase of protease and rnase transformation was recorded
 Incase of dnase no transformation is recorded

Avery et al (1944) revealed the chemical nature of the transforming

substance to be DNA. With the help of experiment they showed that
DNA isolated from SIII strained bacteria could exchange the pathogenic
properties to RII strain bacteria.

Two conclusions were derived….

1- Active factor is DNA which can cause transformation

2- SIII factor contain active factor

EXTRA:-

1- Remove the lipids and carbohydrates from a solution of heat

killed S-cells. Proteins, RNA and DNA remains.
2- Subject the solution to treatment of enzymes to destroy either
the proteins. RNA or DNA.
 3- Add a small portion of each sample to a culture containing R cells.
Observe wheather trasformation occurred by testing for the
prescence of virulent S cells.

DNA REPLICATION

 Self synthesis of DNA molecule

 Occurs only durin S-phase of cell cycle
 Molecule of DNA which is replicated is called Parent DNA
 Molecules produced in this pricess is called daughter DNA
 Parent DNA molecule after replication give rise two daughter
DNA molecules.

SEMI CONSERVATIVE MODEL:-

 Model was given by Watson and Crick, who also proposed

structure of DNA
 According to this model, the parent DNA molecule become
unwind and lose its base pairs (unzipping)
 CENTRAL DOGMA
It provides the basic framework for how genetic information
flows from a DNA sequence to a protein product inside cells.
This process of genetic information flowing from DNA to
RNA to protein is called gene expression.

OKAZAKI’S FRAGMENT
Okazaki fragments are short sequences of DNA
nucleotides (approximately 150 to 200 base pairs long in
eukaryotes) which are synthesized discontinuously and later
linked together by the enzyme DNA ligase to create the
lagging strand during DNA replication.
 GENE EXPRESSION:-
DEFINITION:-

The mechanism in which the genatic information that resides in DNA,

first flows down into the mRNA by the process of transcription and then
by the process of translation.

EXPLANATION:-

The instructions to make proteins are contained in our DNA.

 DNA contains gene. A gene is continous string of nucleotides

containing a region that codes for RNA molecule.
 This region begin with a promoter and ends in a terminator.

PROMOTER:-

Promoter is regulatory region of the gene

which provides binding sites for RNA
polymerase.

 It is present towards the 5’ end of

coding strand.

 In prokaryotes, there are two bonding sites are located on

promoter i.e.
1. TATAAT also called -10 sequence
2. TTGACA also called -35 sequence
 In eukaryotes, also two bonding sites…
1. TATA (TATA box) also called -25 sequence
2. CAAT (CAAT box) also called -70 sequence
  Name of this sequences (-10, -35, -25 and -70) refer to psition that
these sequences are located before the initiation site of structural
region of the gene.

 Genes also contained regulatory sequences that can be found

near the promoter ar at more distant location.
 For some genes, the encoded RNA used to synthesize a protein, in
a process called gene expression.
 For this gene expression can be divided into two process..
1. Transcription
2. Translation

TRANSCRIPTION:-

DEFINITION:-

Transcription is the first step in gene expression. It involves copying a

gene's DNA sequence to make an RNA molecule. Transcription is
performed by enzymes called RNA polymerases, which link nucleotides
to form an RNA strand (using a DNA strand as a template).

EXPLANATION:-
OCCURANCE:-

In eukaryotic cells transcription occurs in nucleus, where DNA is used as

template to make mRNA.

RNA POLYMERASE ENZYME:-

RNA polymerase consists of four subunits…

1. Alpha
2. Beta
3. Beta’
4. Sigma
 Only the first three
subunits are
required for
polymerase activity
and considered the
core enzymes
(needed for catalytic
activity).
 While the sigma factor is required for RNA polymerase to bind to
the promoter.
 It similar to the DNA polymerase in that it also adds nucleotides to
the 3’ end of the growing polypeptide chain but unlike DNA
polymerase it does not require primer to perform polymerase
activity.
 In prokaryotes, only one type of RNA polymerase is found.
 While in eukaryotes, there are three types of RNA namely..
 1. RNA polymerase I
2. RNA polymerase II
3. RNA polymerase III
 RNA polymerase I sythesize ribosomal RNA.
 RNA polymerase II synthesize messenger RNA
 RNA polymerase III synthesize Transfer RNA

MECHANISM:-

During transcription, the DNA in the gene is used as a template to

make a messenger RNA strand with the help of the enzyme RNA
polymerase to bind. This process occurs in three stages….

1. Initiation stage
2. Elongation stage
3. Termination stage

INITIATION STAGE:-

During initiation , the promoter region of the gene function as

recognition site for DNA polymerase to bind.

RECOGNITION SEQUENCE or SITE or REGION:-

A regulatory sequence is a segment of nucleic acid which is capable of

increasing or decreasing the expression of the specific genes within
organisms.
  This is where the majority of gene expression is controlled by
either permitting or blocking access to this site by RNA
polymerase.
 Binding of RNA polymerase causes the DNA double heliix unwind ,
base pairs are broken down , and a bubble like structure, the
transcription bubble is appeared.

ELONGATION:-

 As the RNA polymerase binds to promoter, sigma factor is

released and remaining core enzymes extends the polymerization
of ribonucleoside triphosphates (rNTP).
 It does not require primer to initiate polymerization.
 One of the two strand of the gene acts as template for
transcription.

ANTISENSE or NON CODING:-

This template strand is also called antisense because mRNA is

complementary to this strand.

CODING or SENSE STRAND:-

 The other strand of the gene is called coding or sense strand.

MECHANISM:-

 In elongation stage RNA polymerase keep on moving from 5’ to 3’

direction towards the terminator region, beside it transcription
bubble also moves along the DNA, leaving the growing RNA strand
protruding (stick out) from the bubble.
 As the complementary bases pair up, the DNA polymerase links
nucleotides to the 3’ end of the growing messenger RNA
molecule.
 Once the RNA polymerase reaches the terminator portion of the
gene, the messenger RNA transcripts is complete and the RNA
polymerase, the DNA strand and the mRNA transcript dissociate
from each other.

TERMINATION STAGE:-

The sequence of the terminator region stop the synthesis of mRNA.

 The terminator region consists of a series of GC base pairs

followed by series of AT base pairs.

GC hairpin:-
 The part of mRNA which is transcribed in this region, project form a
loop likes structure called GC hairpin followed by a small tail of AU
nucleotides.

 The GC hairpin causes the RNA polymerase to stop the synthesis

of mRNA.