Difference between revisions of "Part:BBa K3093000:Design"
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===Design Notes=== | ===Design Notes=== | ||
ChbR is encoded within the chb operon itself. In the presence of chitobiose, ChbR binds to two direct repeats located between two NagC binding sites and activates the transcription of the chb operon by overlapping the -35 box of chbBp. In the absence of chitobiose, ChbR acts as a repressor of the chb operon. Monoacetylchitobiose-6-P has been experimentally determined to be the inducer for ChbR. | ChbR is encoded within the chb operon itself. In the presence of chitobiose, ChbR binds to two direct repeats located between two NagC binding sites and activates the transcription of the chb operon by overlapping the -35 box of chbBp. In the absence of chitobiose, ChbR acts as a repressor of the chb operon. Monoacetylchitobiose-6-P has been experimentally determined to be the inducer for ChbR. | ||
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The sequence of chbR was obtained from E.coli K12 MG1655 via PCR.Then we performed inverse PCR to achieve the site-directed mutation of amino acids of chbR. According to the literature, we chose three site-directed mutation: chbRN238S | The sequence of chbR was obtained from E.coli K12 MG1655 via PCR.Then we performed inverse PCR to achieve the site-directed mutation of amino acids of chbR. According to the literature, we chose three site-directed mutation: chbRN238S | ||
Revision as of 15:41, 16 October 2019
chbR N238S Y30C
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
ChbR is encoded within the chb operon itself. In the presence of chitobiose, ChbR binds to two direct repeats located between two NagC binding sites and activates the transcription of the chb operon by overlapping the -35 box of chbBp. In the absence of chitobiose, ChbR acts as a repressor of the chb operon. Monoacetylchitobiose-6-P has been experimentally determined to be the inducer for ChbR.
The sequence of chbR was obtained from E.coli K12 MG1655 via PCR.Then we performed inverse PCR to achieve the site-directed mutation of amino acids of chbR. According to the literature, we chose three site-directed mutation: chbRN238S
Source
ChbR was obtained from the genomic sequence of Escherichia coli str. K-12 substr.MG1655
References
[1]Plumbridge J, Pellegrini O. Expression of the chitobiose operon of Escherichia coli is regulated by three transcription factors: NagC, ChbR and CAP. Mol Microbiol. 2004 Apr;52(2):437-49. PubMed PMID: 15066032.