Lecture 1 – Recombinant DNA
Recombinant DNA technology is used in all aspects of biology:
Medicine: a) Insulin,
b) HGHc) Vaccinesd) Factor 8
e) Forensic DNA fingerprinting,f) Gene therapyg) Disease diagnosis
Lecture 1 – Recombinant DNA
a) Insulin,b) HGHc) Vaccinesd) Factor 8
Medicine:
Lecture 1 – Recombinant DNA
Forensic DNA fingerprinting:
Lecture 1 – Recombinant DNA
Production of valuablepharmaceuticals in transgenic sheepmilk:
Lecture 1 – Recombinant DNA
Human genome project:
Figure 20.0 DNA sequencers
Lecture 1 – Recombinant DNA
Agriculture : Development of novel varieties of plants :
a) Increased protein and yield b) Increased vitamin contentc) Pest and disease resistance.
Lecture 1 – Recombinant DNA
Agriculture : Development of novel varieties of plants
a) Increased protein and yield b) Increased vitamin contentc) Pest and disease resistance.
Figure 20.20 “Golden” rice contrasted with ordinary rice
“Golden rice” with increasedlevels of vitamin A
Lecture 1 – Recombinant DNA
Lecture 1 – Recombinant DNA
This technology is called:
Recombinant DNA,
Gene Cloning,
Genetic Engineering,
Transgenic technology
Ability to take a gene from one organism, and stably insert it into another with a view to expression
Lecture 1 – Recombinant DNA
Recombinant DNA technology involves:
1) A method of cutting DNA molecules
2) A method of joining DNA molecules
3) A method of introducing DNA molecules into cells and organisms
4) A method of identifying recombinants
5) A method to drive expression of the foreign gene in an alien environment.
2) A method for isolating DNA molecules
Lecture 1 – Recombinant DNA
1) A method of cutting DNA molecules – Restriction Enzymes.
Restriction enzymes are bacterial enzymes which have evolved to protectBacteria from phage attack.
Incoming phage DNA is cleaved, but bacterial DNA is not because it is methylated.
Lecture 1 – Recombinant DNA
Restriction enzymes are named after the bacterium from which they were first isolated:
Eco R1 – Escherichia coli – G A A T T C 6 bp
Bam H1 – Bacillus amyloliquefaciens – G G A T C C 6 bp
Sau 3A – Staphylococcus aureus – G A T C 4 bp
Not 1 – Nocardia – - G C G G C C G C 8 bp
Hind 3 – Haemophilus influenzae – A A G C T T 6 bp.
Lecture 1 – Recombinant DNA
Restriction enzymes cleave DNA at SPECIFIC sequences:
To give Blunt (flush) ends and Sticky (cohesive) ends