Part:BBa_K4645012:Design

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.