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 (CaCl2)

• Marker• Cloning Site

LB Broth and Agar

• Lysogeny Broth– Luria Broth– Luria-Bertani

• Nutritionally Rich– Peptides, casein– Vitamins– Minerals (NaCl)– Trace elements

• Biotech– Cultivate E.coli

Procedure• Follow lab manual!!• Reader

– Directs– Keeps time

• Materials– LB plates

• 2 types– LB– LB-amp

• 2 situations– Before– After

• Label, Label, Label– pGreen plasmid– CaCl2– 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