1 2004 Biology Olympiad Preparation Program 2 BIOTECHNOLOGY 2004 Biology Olympiad Preparation Program 3 RECOMBINANT DNA TECHNOLOGY Recombinant DNA technology involves sticking together bits of DNA from different sources. Made possible because DNA & the genetic code are universal. 2004 Biology Olympiad Preparation Program 4 Restriction endonucleases Bacteria use restriction enzymes to cut foreign DNA. Restriction enzymes are specific for a particular restriction site. Restriction fragments are produced by digesting DNA with a particular restriction enzyme. EcoRI – E scherichia co li, strain R , enzyme 1 . AluI 2004 Biology Olympiad Preparation Program 5 Sticky & blunt ends - C T T A A - G G - A A T T C - G - G C A T C - - C T T A A - G - A G - T C C T – G A - T T – A A - - A G - T C Sticky ends must be compatible for complementary base pairing. Blunt ends do not have this requirement. 2004 Biology Olympiad Preparation Program 6 Gene cloning – an overview Isolate plasmid DNA from bacteria Cut with RE Isolate human DNA with gene of interest Cut with same RE Grow bacteria Isolate bacterial clone carrying gene of interest Mix & ligate fragments with DNA ligase Transform bacteria 2004 Biology Olympiad Preparation Program 7 DNA isolation & restriction amp R lacZ Restriction site Cut with RE Gene of interest Cut with same RE
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BIOTECHNOLOGY RECOMBINANT DNA TECHNOLOGYmembers.optusnet.com.au/~jbalasuriya/Notes/Lecture 11.pdf · RECOMBINANT DNA TECHNOLOGY Recombinant DNA technology involves sticking together
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2004 Biology Olympiad Preparation Program 2
BIOTECHNOLOGY
2004 Biology Olympiad Preparation Program 3
RECOMBINANT DNA TECHNOLOGY
Recombinant DNA technology involves sticking together bits of DNA from different sources.
Made possible because DNA & the genetic code are universal.
2004 Biology Olympiad Preparation Program 4
Restriction endonucleases Bacteria use restriction enzymes to cut foreign DNA.
Restriction enzymes are specific for a particular restriction site.
Restriction fragments are produced by digesting DNA with a particular restriction enzyme.