Part:BBa_K2267005
c-di-GMP connected with PAsr
Cyclic dimeric guanosine monophosphate (c-di-GMP)
Cyclic dimeric guanosine monophosphate (c-di-GMP) is a new type of second messenger molecule that is ubiquitous in bacteria. Bacteria utilize it to perceive cell surface signals and activate targets within cells, which involves amplification of the original signal and the subsequent expression of a series of specific genes in the cell. These genes affects the various physiological and biochemical processes in the cells. c-di-GMP control a wide range of process, and has a role in transcription, translation and post-translation levels. In addition, c-di-GMP participates in the regulation of multiple biological functions and affects biofilm formation. In a nutshell, it is a crucial elements to the formation of membrane-like bacterial cellulose and the improvement of BC production.
This experiment attempts to improve BC production by over-expressing PGM and c-di-GMP, a small signal transfer molecule of bacteria. The goal is to implement c-di-GMP to coordinate with PGM and realize the high yield of BC. We have acquired c-di-GMP from genome sequence of G.xylinus via PCR. We have acquired pH-responsive PAsr from the K12 genome sequences of E.coli via PCR. Through PCR overlap extension, we have acquired gene sequences PAsr+c-di-GMP.
Figure 1. c-di-GMP plays a significant role in regulating the biosynthesis of the membrane. The gene circuit is composed of pH-responsive promoter Asr(Part:BBa_K2267000and c-di-GMP with the purpose of overexpressing c-di-GMP
Expressed
Figure 2. Agarose Gel Electrophoresis for c-di-GMP DNA
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 367
Illegal AgeI site found at 130 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 457
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