Cloning of the genetic components involved in cAMP uptake in Escherichia coli
碩士 === 國立交通大學 === 生化工程研究所 === 94 === Cyclic adenosine monophosphate（cAMP）is known to be the important regulator in both prokaryotes and eukaryotes. Previous studies showed that exogenous cAMP could recover the phenomenon caused by cAMP mutation, and proposed that the uptake of cAMP into E.coli was p...
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| Format: | Others |
| Language: | zh-TW |
| Published: |
2005
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| Online Access: | http://ndltd.ncl.edu.tw/handle/91121119761367476808 |
| Summary: | 碩士 === 國立交通大學 === 生化工程研究所 === 94 === Cyclic adenosine monophosphate（cAMP）is known to be the important regulator in both prokaryotes and eukaryotes. Previous studies showed that exogenous cAMP could recover the phenomenon caused by cAMP mutation, and proposed that the uptake of cAMP into E.coli was performed by facilitated diffusion process. However, the mechanism is still unclear. To investigate whether any other molecules take part in the process of cAMP import and the functions of the molecules, mutant library was established by sending transposon Tn5 into E.coli K12 cya mutant strain by electroporation.
Selecting candidate mutant strains by indirect（MacConkey agar、succinate and PCR of crp gene）and direct methods（measurements of intracellular cAMP after adding exogenous cAMP）. 15 mutant clones were analyzed. The results indicated that the changes of membrane structures、 capsules、secretion of membrane protein、some regulator genes and some genes involved in metabolism may interfere with the process of cAMP import. Besides, c-di-GMP and AI-2 may also play the roles in the process of cAMP uptake. These findings need further studies to be confirmed.
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