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General Biology

Chapter 12

Applications of Biotechnology
Applications of
Biotechnology

Biotechnology

the manipulation of organisms or their components to make useful products

dates back to the dawn of civilization when people used yeast to make bread and beer and selectively bred livestock.

today it means the use of DNA technology, modern laboratory techniques for studying and manipulating genetic material.

Biotechnology

Using the methods of DNA technology, scientists

can modify specific genes and move them

between organisms as different as bacteria, plants, and animals.

Organisms that have acquired one or more genes by artificial means are called genetically modified

(GM) organisms.

If the newly acquired gene is from another organism, typically of another species, the

recombinant organism is called a transgenic organism.

Biotechnology

Scientists construct recombinant DNA by combining pieces of DNA from two different sourcesoften from different speciesto

form a single DNA molecule.

Recombinant DNA technology is widely used in genetic engineering, the direct manipulation of genes for practical purposes.

Biotechnology

Scientists have genetically engineered bacteria to mass-produce a variety of useful chemicals, from cancer drugs to pesticides,

and

transferred genes from bacteria to plants and from one animal species to another.

Topics to be Covered

DNA Restriction Enzymes

DNA Gel Electrophoresis

Polymerase Chain Reaction (PCR)

DNA Fingerprinting

Gene Cloning

Genetically Modified Organisms (GMOs)

Reproductive Cloning and Stem Cells

Topic 1: DNA Restriction Enzymes

Topic 1: DNA Restriction Enzymes
Restriction Endonucleases
Restriction Endonucleases

“Murder for a jar of red rum” “God saw I was dog”

EcoRI site:

5’ 3’ 5’
5’
3’
5’
a jar of red rum” “God saw I was dog” EcoRI site: 5’ 3’ 5’ 5’…GAATTC…3’

5’…GAATTC…3’

3’…CTTAAG…5’

a jar of red rum” “God saw I was dog” EcoRI site: 5’ 3’ 5’ 5’…GAATTC…3’

3’

The enzymes generally recognize 4-6 selecting sites for cleavage bases in

The

enzymes

generally

recognize

4-6

selecting sites for cleavage

bases

in

What’s there in a name?

Named for

Bacterial genus,

species,

Strain, and Type

Eg. Eco R1, Bam H1

in a name? Named for • Bacterial genus, • species , • Strain , and •

Why do Bacteria need a Restriction Enzyme

Why do Bacteria need a Restriction Enzyme When they catch Cold

When they catch Cold

How is the bacterial genome safe from its own restriction enzymes?

Methylation of Bacterial Genome
Methylation of Bacterial Genome
Methylation of Bacterial Genome

Classification of Restriction Enzymes

Based upon activity

Type I Type II Type III

Based upon type of free ends

produced

Sticky ends Blunt ends

Sticky Ends
Sticky Ends
Sticky Ends
Blunt Ends
Blunt Ends
Blunt Ends

DNA Ligase

DNA Ligase

Applications of restriction enzymes

Isolated and used for digesting/ cutting DNA molecules of interest for

Generating recombinant DNA molecules =

by joining two DNA different DNA cut using same RE

RFLP = restriction fragment length polymorphism

DNA fingerprinting

Topic 2: Electrophoresis

Movement of charged molecules in electric field.

Molecules move in a matrix of Agarose and get separated as per their size/ molecular weight

DNA/ RNA Gel Electrophoresis

Protein

DNA Gel Electrophoresis

DNA Gel Electrophoresis
DNA Gel Electrophoresis

Gel Electrophoresis

Gel Electrophoresis b.

b.

Gel Electrophoresis b.

Which DNA would Migrate Faster

Super-coiled

Which DNA would Migrate Faster Super-coiled Linear

Linear

Which DNA would Migrate Faster Super-coiled Linear

Visualizing DNA

Electrophoresis in the Laboratory

Visualizing DNA Electrophoresis in the Laboratory
Visualizing DNA Electrophoresis in the Laboratory

End of Lecture