Part:BBa_K3731003
ppk1-vgb-mazE
This part is composed of ppk1, vgb and mazE. From the promoter to the terminator, it express the ppk1, vgb and mazE in sequence. The ppk1 codes PPK1, which can promote the synthesis(major function) and decomposition(minor function) of polyP with the residue of ATP. The vgb can produce HGB so that we can increase the bacteria’s capacity of carrying oxygen, which is beneficial to the growth of bacteria. The mazE codes a kind of antitoxin protein, which can help the bacteria grow longer and produce more polyP.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
iGEM2021_Nanjing China Experiment
This year our team develops s polyphosphate kinase (PPK1) overexpression strategy for achieving maximum intracellular polyphosphate accumulation.Through adding vgb and mazE into the E.coli genome, bacteria grows longer and can survive more severe environment, thus increasing the production of polyP.
We used spectrophotometer to measure OD600 in different culture time, which is an index to estimate the number of bacteria in culture mediums. Then we drew the curve of the number of bacteria against culture time.
We used spectrophotometer to measure OD700 in different culture time, which is an index to estimate the residual inorganic phosphorus content in culture mediums. Then we drew the curve of bacterial function against culture time to measure the polyp production capacity.
Compared with existing literature, we were glad to see an increase in polyP production, which proved an improvement on previous production methods. The success of our experiment inspired team members to make more efforts in the following experiments.
Reference: Kyunghye Ahn and Arthur Kornberg. Polyphosphate Kinase from Escherichia coli[J]. The Journal of Biological Chemistry, 1990, 265, 20, 11734-11739.
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