Difference between revisions of "Part:BBa K3924023"
 
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==Design and Construction==
 
==Design and Construction==
 
We introduce mchE gene from E.coli CA46 into E.coli Nissle 1917. The sequence was obtained from E. coli CA46 microcin operon(GenBank: AJ515251.1). The mchE gene is added into lac operator(on RGP plasmid) so that the mchE 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.<br/>
 
We introduce mchE gene from E.coli CA46 into E.coli Nissle 1917. The sequence was obtained from E. coli CA46 microcin operon(GenBank: AJ515251.1). The mchE gene is added into lac operator(on RGP plasmid) so that the mchE 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.<br/>
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[[Image: mchE.png|center|600px|thumb|'''Figure 1: The design of mchE''']]
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[[Image:T--Tsinghua--part_mchE_construct.png|center|600px|thumb|'''Figure 1: The design of mchE''']]
  
  

Latest revision as of 07:29, 21 October 2021


mchE. mchE MchE transports mature MccH47 to the extracellular side


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 610
    Illegal PstI site found at 823
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 610
    Illegal PstI site found at 823
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 237
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 610
    Illegal PstI site found at 823
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 610
    Illegal PstI site found at 823
  • 1000
    COMPATIBLE WITH RFC[1000]

Profile

Name: mchE
Base Pairs: 1242bp
Origin: Escherichia coli, synthesis
Properties: Microcin H47(MccH47) transport protein

Usage and Biology

mchE was obtained from E. coli strain CA46. It encodes MchE which transports mature MccH47 to the extracellular side.[1]

Design and Construction

We introduce mchE gene from E.coli CA46 into E.coli Nissle 1917. The sequence was obtained from E. coli CA46 microcin operon(GenBank: AJ515251.1). The mchE gene is added into lac operator(on RGP plasmid) so that the mchE 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 mchE


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.