Part:BBa_K778002:Design
fhlA-E363G (engineered trascriptional activator fhlA with a mutation)
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2096
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1375
- 1000COMPATIBLE 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
References
(1)Protein Engineering of the Transcriptional Activator FhlA To Enhance Hydrogen Production in Escherichia coli(Viviana et.al.2009)