Part:BBa_K3598048
AOX1 Promoter_α factor secretion signal_CEN1HC-Br_AOX 1 Terminator
The circuit we transformed into Pichia Pastoris to produce AMP CEN1HC-Br.
Part demonstration
The AOX1-CEN1HC-Br is a composite part consisting of an AOX1 promoter, a CEN1HC-Br coding sequence, and an AOX1 terminator. It is designed for the production of our AMP CEN1HC-Br. We inserted the sequence of the system onto vector pPIC9K and transferred the plasmid into Pichia pastoris GS115 genome. The recombinant strain was cultured for CEN1HC-Br production.
Experiments and Results
We tested the antimicrobial potency of CEN1HC-Br as fermentation product of our Pichia pastoris by adding its solution to plates inoculated with P. acnes and E. coli MG1655. It can be inferred from the results that the fermentation product's effect of killing E. coli MG1655 and P. acnes are not significant. Only a certain amount of bacteria within contact of the products were eliminated and the inhibition zones are quite unclear due to low volume of AMPs dissolved in solutions.
Then, we verified CEN1HC-Br's ability as a fermentation product to kill E. coli MG1655 and P. acnes by adding its solution to liquid culture inoculated with the two bacteria. We set the AMP concentration gradient as 2.5% and 25% in verification with E. coli and 25% with P. acnes because 2.5% proved in prior verification to be too low a concentration for the effects to be significant, and measured the OD600 of the bacteria. The results show that the fermentation product is effective in killing E. coli MG1655 and P. acnes.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 1311
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 937
Illegal XhoI site found at 1191 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1377
- 1000COMPATIBLE WITH RFC[1000]
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