Difference between revisions of "Part:BBa K3924018"

 
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<partinfo>BBa_K3924018 short</partinfo>
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3924018 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3924018 SequenceAndFeatures</partinfo>
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==Profile==
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Name: mchB<br/>
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Base Pairs: 228bp<br/>
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Origin: Escherichia coli, synthesis<br/>
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Properties: Microcin H47(MccH47) precursor<br/>
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==Usage and Biology==
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mchB was obtained from E. coli strain CA46. It encodes the precursor of MccH47. After modified by McmK, McmL, mchC and mchD, the precursor become mature MccH47. The it will be exported by MchE and MchF to the extracellular side.<sup>[1]</sup> <br/>
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==Design and Construction==
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We introduce mchB gene from E.coli CA46 into E.coli Nissle 1917. The sequence was obtained from E. coli CA46 microcin operon(GenBank: AJ515251.1). The mchB gene is added into lac operator(on RGP plasmid) so that the mchB 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: T--Tsinghua--part_mchB_construct.png|center|600px|thumb|'''Figure 1: The design of mchB''']]
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==Reference==
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[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. <br/>
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Latest revision as of 07:17, 21 October 2021


mchB.Microcin H47(MccH47) precursor


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 74
    Illegal PstI site found at 83
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 74
    Illegal PstI site found at 83
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 74
    Illegal PstI site found at 83
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 74
    Illegal PstI site found at 83
  • 1000
    COMPATIBLE WITH RFC[1000]

Profile

Name: mchB
Base Pairs: 228bp
Origin: Escherichia coli, synthesis
Properties: Microcin H47(MccH47) precursor

Usage and Biology

mchB was obtained from E. coli strain CA46. It encodes the precursor of MccH47. After modified by McmK, McmL, mchC and mchD, the precursor become mature MccH47. The it will be exported by MchE and MchF to the extracellular side.[1]

Design and Construction

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


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.