Difference between revisions of "Part:BBa K2997003"

Revision as of 19:12, 19 October 2019

Sigma 54-dependent transcriptional regulator (pchR)

The pchR gene can be translated into activator proteins that can bind with the inducer (p-Cresol) and leads to the conformation of the protein. In other words, the activator protein can bind with p-Cresol to form a protein-p-Cresol complex that will then bind onto the p-Cresol sensing region located just downstream of the pchR gene. The binding of the protein - p-Cresol complex initiates the expression of the downstream gene, in our case GFP.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1738
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 1151
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 1196
Illegal AgeI site found at 179
Illegal AgeI site found at 1079
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1105

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 <partinfo>BBa_K2997003 short</partinfo>
-The construction and digestion of DNA gel shows that the size of the pchR-GFP composite part was as expected.
+The pchR gene can be translated into activator proteins that can bind with the inducer (<i>p</i>-Cresol) and leads to the conformation of the protein. In other words, the activator protein can bind with <i>p</i>-Cresol to form a protein-<i>p</i>-Cresol complex that will then bind onto the p-Cresol sensing region located just downstream of the pchR gene. The binding of the protein - <i>p</i>-Cresol complex initiates the expression of the downstream gene, in our case GFP.
-The pchR-GFP composite part (BBa_K2997008) was constructed from IDT DNA synthesis. After undergoing a few rounds of overlap PCR to combine 3 separate fragments into one long composite part, the part is then inserted into pSB1C3 and electroporated into E. coli Top 10. After allowing the transformed E. coli Top 10 to grow on LB+Cm plates, colony PCR and sequencing was carried out to determine the clone we got was positive or not. The colony PCR results indicated positive results, but the sequencing results indicated that random mutations have occurred on the pchR-GFP composite part.