1. Bacterial Genetics & Transformation
Preparation for the pGlo Lab

2. Bacterial Genome
Circular
Little protein
Condensed DNA = nucleoid

3. 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)

4. Genetic Recombination
Combining DNA from two sources
Usually two bacterial cells or bacteria & a phage

5. Conjugation & Plasmids
Conjugation: Direct transfer of genetic material b/w bacterial cells via a sex pilus that attaches to the recipient

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

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

8. 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

9. 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

10. 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).

11. Expression of Green Fluorsecent Protein

12. Predict Which Plates Will Glow