1. Prodigiosin Production in E. Coli
Brian Hovey and Stephanie Vondrak

2. What is Prodigiosin?
A secondary metabolite of various strains of Serratia, and other Gram negative gammaproteobacteria.
It is responsible for the red pigment produced by Serratia marcescens.
Produced under the control of 14 genes(pigA-pigN)

3. S. marcescens
S. marcescens is a species of Gram negative, rod shaped bacteria
Known to cause many nosocomial infections
Thrives in high moisture environments

4. Significance?
Recently, has gotten attention for its newfound benefits.
Such as: antibacterial, antifungal, antiprotozoal, antimalarial, immunosuppressive, and anticancer properties
Has no or little toxicity to cell lines (may operate as a cell cycle regulator)

5. pigI Gene
We chose pigI because it is involved in one of the beginning pathways of MBC(4- methoxy-2,2`-bipyrrole-5-carbaldehydе)
This is a precursor of prodigiosin

6. Prodigiosin Pathway
G.O.I.

7. How
We located the gene sequence in NCBI, with the accession number: AJ833002, and has 1473 base pairs.
We will amplify the gene by PCR with primers (TBD)
Amplification will be checked by gel electrophoresis
Biobrick attachment
Using BBa_I Tet repressor generator.
We will choose a plasmid with specific antibiotic resistances
Some strains of S. marcescens are known to be resistant to a number of antibiotics naturally
The restriction site will be cut by TBD restriction enzyme

8. How, continued Hybrid DNA will be inserted into plasmid, then ligated.
Plasmid will then be inserted into host bacteria (E. coli)
Detection of pigI product will be determined by SDS-PAGE
As a backup, we will use the Tetracycline repression Biobrick to test for pickup of the target gene
If resistant to (TBD), then it is assumed that the plasmid was successfully picked up, those cultures can then be tested for resistance to tetracycline; if the bacteria die, the Biobrick was successful and it is assumed that the gene also was.

9. References
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