Coding

Part:BBa_K4645012:Design

Designed by: Yuanpu Zhang   Group: iGEM23_HZAU-China   (2023-10-11)


The RstAB system is a bacterial two-component regulatory system consisting of the membrane sensor


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 324
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1501


Design Notes

The RstA/RstB system is a bacterial two-component regulatory system consisting of the membrane sensor, RstB and its cognate response regulator RstA. The RstA consists of an N-terminal receiver domain (RD, residues 1–119) and a C-terminal DNA-binding domain (DBD, residues 130–236). Phosphorylation of RstA induces dimerization, which allows two RstA DBDs to bind to a tandem repeat, called the RstA box, and regulate the expression of downstream genes [1]. In the sequence of the acid-sensitive promoter asr, there is a RstA box [2].

Source

The team obtained the RstAB fragment from the E. coli MG1655 genome by PCR using a high-fidelity DNA polymerase. 

Reference

[1]Li YC, Chang CK, Chang CF, Cheng YH, Fang PJ, Yu T, Chen SC, Li YC, Hsiao CD, Huang TH.Structural dynamics of the two-component response regulator RstA in recognition of promoter DNA element. Nucleic Acids Res. 2014 Jul;42(13):8777-88.

[2]Ogasawara H, Hasegawa A, Kanda E, Miki T, Yamamoto K, Ishihama A. Genomic SELEX search for target promoters under the control of the PhoQP-RstBA signal relay cascade. J Bacteriol. 2007 Jul;189(13):4791-9.