Part:BBa_K4242000
FleQ
coding region for FleQ protein, which is a transcription factor that can be combine with FleN and c-di-GMP to regulate the gene transcription under the promoter Ppel.
Usage and Biology
FleQ can control flagella biosynthesis and exopolysaccharide (EPS). scientists found FleQ can activate the expression of flagella biosynthesis genes but inhibit the expression of exopolysaccharide genes. It has an important Usage we can regulate FleQ to control biofilm information and explore medical research.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 868
Illegal XhoI site found at 1321 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 261
Illegal NgoMIV site found at 1269
Illegal NgoMIV site found at 1281
Illegal NgoMIV site found at 1357
Illegal NgoMIV site found at 1442
Illegal AgeI site found at 670 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 283
Illegal SapI.rc site found at 1216
Illegal SapI.rc site found at 1225
FleQ protein bend with DNA
Functional Parameters: CUG_China's contributions
when we constructed the biobrick, we found that there is a PstⅠ enzyme cutting site in its coding sequence. Its coding sequence is not compatible with biobrick assembly, and we cannot directly use it as a part. To make this gene compatible with biobrick, we removed this cutting site by using site-directed mutagenesis. Now the new sequence of fleQ has been registered as a new basic part, and we are happy if this may be helpful to any other future iGEMers.
Blast result of our site-removed FleQ
References
[1]Matsuyama BY, Krasteva PV, Baraquet C, Harwood CS, Sondermann H, Navarro MV. Mechanistic insights into c-di-GMP-dependent control of the biofilm regulator FleQ from Pseudomonas aeruginosa. Proc Natl Acad Sci U S A. 2016 Jan 12;113(2):E209-18.
[2]Hickman JW, Harwood CS. Identification of FleQ from Pseudomonas aeruginosa as a c-di-GMP-responsive transcription factor. Mol Microbiol. 2008 Jul;69(2):376-89.
[3]Baraquet C, Murakami K, Parsek MR, Harwood CS. The FleQ protein from Pseudomonas aeruginosa functions as both a repressor and an activator to control gene expression from the pel operon promoter in response to c-di-GMP. Nucleic Acids Res. 2012 Aug;40(15):7207-18.
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