Coding

Part:BBa_K3924017

Designed by: Wenda Zhang   Group: iGEM21_Tsinghua   (2021-09-30)
Revision as of 07:13, 21 October 2021 by Songwh19 (Talk | contribs) (→‎Profile)

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mchI. It is immune gene that encodes a immunity protein MchI against MccH47


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 99
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 99
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 99
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 99
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 99
  • 1000
    COMPATIBLE WITH RFC[1000]

Profile

Name: mchI
Base Pairs: 210bp
Origin: Escherichia coli, synthesis
Properties: Microcin H47(MccH47) immunity protein

Usage and Biology

mchI was obtained from E. coli strain CA46. It is immune gene that encodes a immunity protein MchI against MccH47.[1]

Design and Construction

We introduce mchI gene from E.coli CA46 into E.coli Nissle 1917. The sequence was obtained from E. coli CA46 microcin operon(GenBank: AJ515251.1). The mchI gene is added into lac operator(on RGP plasmid) so that the MchI can only be expressed induced by IPTG.Due to the time cost of obtaining the sequence, the sequence was submitted to the company for synthesis and we sequenced it to verify the correctness of the synthetic sequence. The sequence was added to RGP plasmid by Hifi DNA assembly replacing the TMAR sequence for further use.

Figure 1: The design of mchI


Reference

[1] Vassiliadis G, Destoumieux-Garzón D, Lombard C, Rebuffat S, Peduzzi J. Isolation and characterization of two members of the siderophore-microcin family, microcins M and H47. Antimicrobial Agents and Chemotherapy. 2010 Jan;54(1):288-297. DOI: 10.1128/aac.00744-09. PMID: 19884380; PMCID: PMC2798501.




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