Biochemistry Department.

Course Code :701

Instructor : Dr/Omneya

NA Denaturation

Outlines :A.
B.
C.
D.
E.
F.
G.

Introduction.
What's meant by DNA Denaturation.
Causes of DNA Denaturation.
Characters of Denatured DNA .
Diseases caused by DNA Denaturation.
DNA renaturation.
Application on DNA denaturation and
renaturation.
H. References.
I. Participants.

A. Introduction
-DNA is a double
helix composed of 2
strands running in
opposite directions .
-Each strand composed
of backbone of deoxyribose sugar
molecules linked
together by phosphate
linkage .

Each sugar molecule

covalently linked to one of
four bases
Adenine,Guanine,Cytosine
andThymine.
The two strands connected
together by H-bond.

A.2. Function of DNA.

1. Stores an organism genetic material in
the nuclei.

2. Replicates itself when dividing.

3. Provide code

or template for
particular sequencing of amino acid that
bond together and make protein.

B.DNA Denaturation
Definition

DNA Denaturation is the

separation of a double strand
into two single strands,
which occurs when the
hydrogen bonds between
the strands are broken.

C.Causes of
Denaturation.
Denaturation can occur when
proteins and nucleic acids are
subjected to :
1-elevated temperature.
2-extremes of pH.
3-nonphysiological
concentrations of salt, organic
solvents, urea, or other
chemical agents.

.1-elevated temperature.
The most common
type of denaturation
is thermal
denatuation.

Tm : temp at which
DNA is half
denatured.
*above Tm DNA is
single strands.
*below tm DNA is
double strands.

C.2-extremes of pH
At high PH the
hydroxide
ions (( negatively
charged
ions )) can pull hydrogen
ions from base pairs
forming H bond between
two strands causing
them to separate.

C.3 non physiological salts

Low salt concentration could also denature
DNA double-strands by removing ions that

stabilize the negative charges on the two

strands from each other.

D.

Characters of Denatured DNA

1. Hyperchromic : increase the absorbance

(A260) upon denaturation.
2.

The rate of increase in absorbance is

directly proportional to the rate of
denaturation.

3. Viscosity decrease upon denaturation.

E.Diseases caused by DNA denaturation

DNA repair deficiency disorder is a medical

condition due to reduce functionality of DNA
repair .
DNA repair defects can cause both an
accelerated aging disease and an increase
risk of cancer.
Examples for DNA defects accelerated aging :
1.Bloom syndrome.
2.Fanconi`s anemia.

DNA repair defects and increased

cancer risk :
When a mutation is present in a
DNA repair gene, the repair gene will
either not be expressed or expressed
in an altered form so the repair
function will likely be deficient .
DNA synthesis leading to mutations ,
some of which may give rise to
cancer .

F.DNA Renaturation
Renaturation or annealing :
I.

is the formation of base repairs and

complementary strands of DNA come back
together.

II. Renaturation occurs if double stranded DNA is

heated above tm then the temperature is slowly
decreased under appropriate conditions.

F.Applications on DNA denaturation and renaturation

1. Understanding the genome size

and complexity.
2. Understanding genetic relatedness.
3. Understanding relative proportion
of single copy and repetitive
sequences.

G.References

1.
2.
3.
4.

Lecture notes illustrated

biochemistry I .
Harper`s illustrated
Biochemistry.
Lippincott's illustrated reviews.
Oraby`s illustrated reviews of
biochemistry.