PowerLecture: Chapter 16 Studying and Manipulating Genomes

Genetic Changes _____________________________________________________________________________________________ _________________________________ _______________________________

Discovery of Restriction Enzymes ___________________________________________________________________________________ _________________________________________________

Specificity of Cuts _____________________________________________________ _____________________________________________________________________________________

Making Recombinant DNA 5’ G A A T T C 3’ C T T A A G one DNA fragment another DNA fragment 5’ G A A T T C 3’ 3’ C T T A A G 5’

Making Recombinant DNA nick 5’ G A A T T C 3’ 3’ C T T A A G 5’ nick DNA ligase action G A A T T C C T T A A G

enzyme recognition site A G cut fragments 3’ 5’ enzyme recognition site G C T A another DNA fragment A T C 3’ 5’ G one DNA fragment DNA ligase action nick G C T A 3’ 5’ G C T A 3’ 5’ Stepped Art Fig. 16-2, p.244

Using Plasmids ________________________________________________

Plasmids Fig. 16-3a, p.244

Plasmids Fig. 16-3b, p.244

Restriction enzyme cuts molecule of chromosomal DNA or cDNA fragments with sticky ends Foreign DNA, plasmid DNA, and modification enzymes are mixed recombinant plasmids containing foreign DNA Same enzyme cuts same sequence in plasmid DNA plasmid DNA with sticky ends host cells containing recombinant plasmids Stepped Art Fig. 16-4, p.245

Making cDNA (complimentary DNA) Fig. 16-5, p.245

Gene Libraries ___________________________________________

Using a Probe to Find a Gene ______________________________________________________ ______________________ ____________________________________________________________

Use of a Probe Colonies on plate Cells adhere to filter Cells are lysed; DNA sticks to filter Probe is added Location where probe binds forms dark spot on film, indicates colony with gene

Amplifying DNA __________________________________________________________ _____________________________

Polymerase Chain Reaction _________________________ _____________________________________________ ________________________________________________________________ ________________________________

Polymerase Chain Reaction Double-stranded DNA to copy DNA heated to 90°– 94°C Primers added to base-pair with ends Mixture cooled; base-pairing of primers and ends of DNA strands DNA polymerases assemble new DNA strands Stepped Art Fig. 16-6, p. 256

Polymerase Chain Reaction Mixture heated again; makes all DNA fragments unwind Mixture cooled; base-pairing between primers and ends of single DNA strands DNA polymerase action again doubles number of identical DNA fragments Stepped Art Fig. 16-6, p. 256

Recording the Sequence p.248

Gel Electrophoresis _____________________________ _______________________________________________________________ ________________________________________________

Reaction Mixture ____________________________ __________________

Nucleotides for Sequencing _______________________________ _____________________ _______________________________________________________

Reactions Proceed ____________________________________________________

Recording the Sequence T C C A T G G A C C T C C A T G G A C Recording the Sequence T C C A T G G A T C C A T G G T C C A T G T C C A T T C C A electrophoresis gel T C C __________________ ____________________________________ T C one of the many fragments of DNA migrating through the gel T one of the DNA fragments passing through a laser beam after moving through the gel T C C A T G G A C C A

Recording the Sequence Fig. 16-8b, p.248

DNA Fingerprints ________________________________ ___________________________________________________ ____________________________________________________

Tandem Repeats ________________________________________________________________ ____________________________________________

RFLPs ___________________________________ ______________________________________________________________________ ______________________________________________________________________________________

Gel Electrophoresis Fig. 16-9a, p.249

Gel Electrophoresis Fig. 16-9b, p.249

Analyzing DNA Fingerprints ______________________________________________ _____________________ _______________________________

Genome Sequencing ____________________________________________________________________ _____________________________________________

Genome Sequencing Fig. 16-10a, p.250

Genomics ____________________________________________________________________ _______________________________________________________________________________________

DNA Chips ______________________________________________________________________________________________________ ____________________________________________________________________

DNA Chips Fig. 16-11, p.251

Genetic Engineering ____________________________________________________________________ __________________________________ ________________________

Engineered Proteins _________________________________________________

Cleaning Up the Environment ________________________________________________________________ __________________________________________________________________________________

Can Genetically Engineered Bacteria “Escape”? ________________________________________________________________________________

p.252

Engineered Plants ____________________________________________

Engineered Plants Fig. 16-12a, p.253

Engineered Plants Fig. 16-12b, p.253

The Ti plasmid _________________________________________________________________ ____________________________________________________ plant cell foreign gene in plasmid

The Ti plasmid b The bacterium infects a plant and transfers the Ti plasmid into it. a A bacterial cell contains a Ti plasmid (purple) that has a foreign gene (blue). e Young plants with a fluorescent gene product. c The plant cell divides. d Transgenic plants. Fig. 16-13, p.253

First Engineered Mammals ________________________________________________________________ ________________________________

Transgenic Mice Fig. 16-15, p.254

Designer Cattle ____________________________________________________

Genetically Modified Animals Featherless (created by traditional cross breeding) Fig. 16-14a, p.254

Genetically Modified Animals Mira a transgenic goat that makes a human anticlotting factor Fig. 16-14b, p.254

Genetically Modified Animals Fig. 16-14c, p.254

Xenotransplantation _____________________________________________________

Safety __________________ ____________________________________________________

Safety _________ ________________________________

The Human Genome Initiative Goal - Map the entire human genome ____________________________________________ ____________________________________________________________________ _____________________________________________

Using Human Genes ________________________________________________________________ ________________________________________________________________________________________________ ________________________________________________

Ethical Issues ________________________________________________________________ _____________________