Bacterial Genetics & Transformation

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Presentation transcript:

Bacterial Genetics & Transformation Preparation for the pGlo Lab

Bacterial Genome Circular Little protein Condensed DNA = nucleoid

Mutation as a Source of Genetic Variation Bacteria reproduce asexually by binary fission Mutation can increase genetic diversity when reproduction rates are high b/c of short generations (fitness)

Genetic Recombination Combining DNA from two sources Usually two bacterial cells or bacteria & a phage http://www.youtube.com/watch?v=MRBdbKFisgI

Conjugation & Plasmids Conjugation: Direct transfer of genetic material b/w bacterial cells via a sex pilus that attaches to the recipient http://www.biologie.uni-hamburg.de/b-online/library/micro229/terry/images/anim/Fmating.gif

Plasmids Small circular piece of DNA, separate from bacterial chromosome can be reversably integrated into the bacteria’s chromosome Confer favorable traits

pGLO Bacterial Transformation Lab Transform e. coli bacteria Move genes from one organism to another with the aid of a plasmid

P GLO Plasmid pGLO plasmid encodes the gene for GFP and a gene for resistance to the antibiotic ampicillin pGLO gene is turned on (regulated) in the presence of the sugar, arabinose

Gene Regulation Not all genes are “on” all the time—why? Promoter: Located on the DNA template where RNA polymerase begins transcription of Mrna Operon: A cluster of genes controlled by a single promoter

Arabinose Operon Bacteria produce three enzymes (araB, araA and araD) to break down arabinose sugar The “ara” genes are clustered together and controlled by a single promoter (Pbad).

Expression of Green Fluorsecent Protein

Predict Which Plates Will Glow