Signalling

Part:BBa_K1897001:Design

Designed by: Choi Yan Ru   Group: iGEM16_NUS_Singapore   (2016-10-09)
Revision as of 10:09, 10 October 2016 by Cyr95 (Talk | contribs) (→‎Source)


HasA hemophore expression unit


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 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 480


Design Notes

The presence of a hexa histidine tag would allow for easy protein purification and detection using anti-his antibodies. The promoter BBa_K525998 was chosen as we wished to induce high levels of HasA for purification thus we required a T7 promoter that could be used to induce expression at high levels in E. coli strain BL21(DE3). The terminator BBa_B0015 was chosen as it is commonly used and has a high efficacy.


Source

The HasA gene is originally found in the Serratia marcescens. We synthesised the part from the Serratia marcescens genome, obtaining the gene sequence from the National Center for Biotechnology Information (NCBI) database (http://www.ncbi.nlm.nih.gov/nuccore/X81195.2), including a hexa histadine tag before the stop codon. The Promoter, RBS and terminator are biobrick parts that were obtained from the Distribution kits from iGEM.

References

Serratia marcescens hasA, has B, hasD, hasE, hasI, hasR and hasS genes. Retrieved May 2016 from http://www.ncbi.nlm.nih.gov/nuccore/X81195.2

Cescau, S., Cwerman, H., Letoffe, S., Delepelaire, P., Wandersman, C., & Biville, F. (2007). Heme acquisition by hemophores. Biometals, 20(3-4), 603-613.

Wandersman, C., & Delepelaire, P. (2004). Bacterial iron sources: from siderophores to hemophores. Annu. Rev. Microbiol., 58, 611-647.