Introduction to

Biotechnology

Manipulating and Analyzing DNA

Uses of Biotechnology

Analyzing human DNA

Genetic diseases

DNA fingerprinting

Forensics

The Human Genome Project

Identifying all genes in the human genome

Gene therapy

Curing disease

Genetic EngineeringDefinition: introducing a set of genetic information

(genes) directly into an organisms DNA

How is this done?

With use of restriction enzymes

Plasmid DNA

Restriction EnzymesProteins originally

made to protect

against viruses

Restriction Enzymes

chut DNA at specific

sequences called

recognition sites

Recognition sites

are sequences of

DNA that enzymes

recognize

The “Ends”Two types of cuts result from restriction enzymes: “Sticky” ends leave exposed bases ready to hydrogen

bond. Eco RI below is an example.

“Blunt” ends leave NO bases exposed Eco RV below is an example

DNA ligase joins “the ends” of cut DNA together

DNA Recombination

DNA fragments cannot function all by

themselves

They must become part of the genetic

material of LIVING cells before the genes

they contain can be activated

DNA fragments + DNA from a living cell =

Recombinant DNA

DNA fragments are often inserted in

plasmids (found in bacteria) that have

been cut with the same sticky ends

Plasmids

Circular double-stranded DNA (bacterial)

Used in biotechnology because they are

circular, small, and replicate on their own

Plasmids + DNA of interest

Restriction enzyme

EcoRI used to isolate

gene of interest

Same restriction

enzyme EcoRI used

to cut plasmid DNA

Sticky ends match up.

Gene of interest is

inserted into

plasmid=recombinant

DNA

Genetic Engineering Example

Insulin is made for diabetics

a) Cut open plasmid and DNA with gene encoding for insulin with restriction enzymes

b) Mix cut plasmid with cut DNA (they have the same sticky ends)

c) Seal with ligase

d) Insert recombinant DNA (plasmid) into bacteria

e) Recombinant DNA replicates and bacteria divides

f) DNA is transcribed and translated = insulin

Engineering New Organisms

Transgenic Organisms:

Organisms that contain

“foreign” DNA are termed

transgenic

May be bacteria, plants,

or animals

Examples of GFP gene

This All Involves One Thing…

DNA Sequencing:

reading or identifying

the sequence of

bases along the

length of a DNA

molecule AND

understanding what

that sequence codes

for…