RNA

Part:BBa_K581006

Designed by: Qin Xiao   Group: iGEM11_Peking_Signaling   (2011-09-26)
Revision as of 16:51, 5 October 2011 by Chenmiaomiao (Talk | contribs)

Pc+ptsG(wt)-GFP (ptsG(wt) 5' UTR fused with gfp)


PtsG is a glucose permease which is subordinate to phosphotransferase system serve as a transpotor. Here,we studied this mRNA perform the conjugate part of the small RNA regulator sgrS(wt). ptsG mRNA is regulated by SgrS by short, imperfect base-pairing interactions, and its expression is thus repressed.

Teppei Morita et.al’ s work suggests that two mutations (C85G and C87G) in ptsG mRNA could completely impair the ability of SgrS to downregulate its expression, while compensatory mutations of SgrS (G178C and G176C) restore the gene silencing ability. These results indicate that it is the base pairing of the two RNAs rather than particular nucleotides that is important for SgrS action. They have also illustrated that sequence outside this region, even though complementary, is rather dispensable for the efficient silencing (Kawamoto et al., 2006). This makes mutant ptsG/SgrS pairs orthogonal to genetic context of the host cell. Therefore we choose this couple of conjugate mRNA/sRNA as the foundation of our comparator device design.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 790



[edit]
Categories
//chassis/prokaryote/ecoli
transcriptional
Parameters
biology5' regulatory region of ptsG(wt) is the target of sRNA SgrS(wt)
chassisE. coli DH5α
n/aPc+ptsG(wt)-GFP (ptsG(wt) 5' UTR fused with gfp)