Part:BBa_K822002
Colicin E1 + immunoprotein
Coding sequence for the channel-forming protein Colicin E1 of E. coli and its associated immunoprotein. Colicins are bacteriocins produced by some strains of E. coli, being lethal to other strains of E. coli and some other species. Colicin E1 has been researched for use as food preservatives and in treating disease. This part is derived from BBa_K150009.
Testing
The part was placed under a constitutive promoter and cell growth experiments performed to determine if cell growth would be inhibited in samples treated with lysate of the cells containing the part. Experiments showed a delayed response with differences between those samples treated with lysate from the putative colicin-producing cells and those treated with lysate from a negative control being apparent after about 3 hours.
Cell cultures with plasmids not containing the colicin part were prepared and kept at 37 °C over night. Two new cultures were then prepared using cells from the overnight cultures. Cells containing the colicin part in an ampicillin resistant plasmid were lysed and the resulting lysate added to one of the cell cultures. The two cultures, one containing lysate of presumed colicin producing cells, the other containing lysate from cells not producing colicin, were then kept at 37 °C in a shaking incubator with 2 mL samples being taken out approximately once an hour. The OD of the samples were measured and compared. The OD of the colicin culture grew significantly slower than the negative control culture, indicating constitutive colicin production in this culture as expected. This experiment was carried out twice (the second time with two cultures containing the colicin plasmid) and the results are illustrated in the graphs below.
Safety
Colicin E1 is considered safe for use as a food preservative, and humans are likely to be chronically exposed to low levels of colicins produced by commensal gastrointestinal bacteria. [1] For this reason, we consider this part to present no particular safety issues.
References
[1]: "Applications and Mechanisms of Colicin E1", Brenda Sue Patton, Ph.D. Thesis - Iowa State University, 2007. ISBN 0549357262.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 301
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