General Biology

Chapter 12

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.

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.

Biotechnology

DNA Restriction Enzymes DNA Gel Electrophoresis Polymerase Chain Reaction (PCR) DNA Fingerprinting Gene Cloning Genetically Modified Organisms (GMOs) Reproductive Cloning and Stem Cells

Topics to be Covered

Topic 1: DNA Restriction Enzymes

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

EcoRI site:

Restriction Endonucleases

5GAATTC3 3CTTAAG5

5 3

3 5

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

Named for

Bacterial genus,

species,

Strain, and

Type

Eg. Eco R1,

Bam H1

Whats there in a name?

Why do Bacteria need a Restriction Enzyme

When they catch Cold

How is the bacterial genome safe from its own

restriction enzymes?

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

Blunt Ends

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

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

Topic 2: Electrophoresis

DNA Gel Electrophoresis

b.

Gel Electrophoresis

Which DNA would Migrate Faster

Super-coiled Linear

Electrophoresis in the Laboratory

Visualizing DNA

End of Lecture