Generator

Part:BBa_K778005:Design

Designed by: Yuka Yamazaki   Group: iGEM12_UT-Tokyo   (2012-09-23)

RBS-fhlA363-d.term


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2115
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1394
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Original fhlA gene we cloned from E. coli JM109 has recognition site of PstⅠ.This illegal (inappropriate for biobrick) site was deleted by overlap extension PCR generating mutation (G to A).

Only one mutation (A to G, Thr to Ala) was found at 1906bp in this biobrick part(different from E. coli K-12 MG1655). This may have originated from cloning PCR. It is also possible that this is minor mutation of genome of JM109 we used.

E363G (A to G, glutamic acid to glycine) point mutation was generated by inverce PCR. This mutation increased H2 production under the conditions in the previous research (Viviana et.al.2009(1)). However, in our research ( team UT-Tokyo 2012 [http://2012.igem.org/Team:UT-Tokyo/Project/H2_E.coli] ), this mutation is

Source

biobrick parts

References

(1)Protein Engineering of the Transcriptional Activator FhlA To Enhance Hydrogen Production in Escherichia coli(Viviana et.al.2009)