Coding

Part:BBa_K4242000

Designed by: zhelin Jiang   Group: iGEM22_CUG-China   (2022-07-12)
Revision as of 13:10, 30 September 2022 by Jiangzhelin (Talk | contribs)


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.

FleQ works in this Part


<center>FleQ work mechanism





Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 868
    Illegal XhoI site found at 1321
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE 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
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 283
    Illegal SapI.rc site found at 1216
    Illegal SapI.rc site found at 1225

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

https://parts.igem.org/File:Blast_result_of_our_site-removed_FleQ.png
fig1. Blast Result of the site-directed sequence and original sequence of fleQ, CTG and CTA code the same amino acid, Leu


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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