Molecular Microbiology Genetic Recombination Vertical Binary fission Budding
Molecular Microbiology
Genetic Recombination • Vertical – Binary fission – Budding • Horizontal – Conjugation – Transduction – Transformation
Conjugation
Transduction
Transformation
Griffith Experiment
Biotechnology • Recombinant DNA • Genetic Engineering • Disease treatments – SCID – SSA – CF • Make products – – Vaccines Enzymes Insulin Hormones
Plasmids • DNA • Replicate independently • Genetic Vectors – – Small Stable Marker inserted Gene expression
Background • Donor • Recipient • Competent Cell – Natural • Change cell wall • Change PM – Artificial • Temperature • Salts (Ca. Cl 2) • Marker • Cloning Site
LB Broth and Agar • Lysogeny Broth – Luria-Bertani • Nutritionally Rich – – Peptides, casein Vitamins Minerals (Na. Cl) Trace elements • Biotech – Cultivate E. coli
Procedure • • Follow lab manual!! Reader – Directs – Keeps time Materials – LB plates • 2 types – LB-amp • 2 situations – Before – After • Label, Label – p. Green plasmid – Ca. Cl 2 – Micropipets – Spreader – Loop inoculation Temperature – Incubator – Baths • Ice • Heat
Results • Marker – Fluorescence • Cloning site – R-ampicillin • Colony observation – Natural light – UV light • Calculate transformation efficiency
Questions? Glow Fish
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