Difference between revisions of "Part:BBa K4242000"
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<partinfo>BBa_K4242000 short</partinfo> | <partinfo>BBa_K4242000 short</partinfo> | ||
| − | This part sequence originated from Pseudomonas aeruginosa encodes FelQ protein. FleQ can repress pel gene expression by a simple roadblock mechanism in which FleQ sits at the pelA promoter and prevents RNA polymerase to access DNA. The binding of c-di-GMP to FleQ induces dissociation of FleQ from the pelA promoter allowing the RNA polymerase to access DNA[1]. FleQ is not only a repressor of pel gene expression in the absence of c-di-GMP but also an activator of pel expression in the presence of c-di-GMP, which suggest two FleQ binding sites at the pel promoter are independently dedicated to activation and repression[2]. What’s more, the pelA promoter occurs distortion needs FleQ work with FleN in the presence of ATP. The DNA bending mediated by FleN occurs only when FleQ is bound to both FleQ boxes. The DNA bending stimulated by FleN occurs only when ATP is present and disappears when c-di-GMP is added[3]. | + | This part sequence originated from <i>Pseudomonas aeruginosa</i> encodes FelQ protein. FleQ can repress <i>pel</i> gene expression by a simple roadblock mechanism in which FleQ sits at the <i>pelA</i> promoter and prevents RNA polymerase to access DNA. The binding of c-di-GMP to FleQ induces dissociation of FleQ from the pelA promoter allowing the RNA polymerase to access DNA[1]. FleQ is not only a repressor of <i>pel</i> gene expression in the absence of c-di-GMP but also an activator of pel expression in the presence of c-di-GMP, which suggest two FleQ binding sites at the pel promoter are independently dedicated to activation and repression[2]. What’s more, the <i>pelA</i> promoter occurs distortion needs FleQ work with FleN in the presence of ATP. The DNA bending mediated by FleN occurs only when FleQ is bound to both FleQ boxes. The DNA bending stimulated by FleN occurs only when ATP is present and disappears when c-di-GMP is added[3]. |
Revision as of 13:56, 8 October 2022
FleQ
This part sequence originated from Pseudomonas aeruginosa encodes FelQ protein. FleQ can repress pel gene expression by a simple roadblock mechanism in which FleQ sits at the pelA promoter and prevents RNA polymerase to access DNA. The binding of c-di-GMP to FleQ induces dissociation of FleQ from the pelA promoter allowing the RNA polymerase to access DNA[1]. FleQ is not only a repressor of pel gene expression in the absence of c-di-GMP but also an activator of pel expression in the presence of c-di-GMP, which suggest two FleQ binding sites at the pel promoter are independently dedicated to activation and repression[2]. What’s more, the pelA promoter occurs distortion needs FleQ work with FleN in the presence of ATP. The DNA bending mediated by FleN occurs only when FleQ is bound to both FleQ boxes. The DNA bending stimulated by FleN occurs only when ATP is present and disappears when c-di-GMP is added[3].
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
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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.