Difference between revisions of "Part:BBa K3924024"

 
 
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<partinfo>BBa_K3924024 short</partinfo>
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3924024 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3924024 SequenceAndFeatures</partinfo>
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==Profile==
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Name: mchF<br/>
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Base Pairs: 2097bp<br/>
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Origin: Escherichia coli, synthesis<br/>
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Properties: Microcin H47(MccH47) ABC transporter protein<br/>
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==Usage and Biology==
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mchF was obtained from E. coli strain CA46. It encodes MchF which transports mature MccH47 to the extracellular side.<sup>[1]</sup> <br/>
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==Design and Construction==
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We introduce mchF gene from E.coli CA46 into E.coli Nissle 1917. The sequence was obtained from E. coli CA46 microcin operon(GenBank: AJ515251.1). The mchF gene is added into lac operator(on RGP plasmid) so that the mchF 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_mchF_construct.png|center|600px|thumb|'''Figure 1: The design of mchF''']]
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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 07:32, 21 October 2021


mchF. mchF transports mature MccH47 to the extracellular side


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 448
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 448
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 448
    Illegal BamHI site found at 1115
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 448
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 448
    Illegal AgeI site found at 1496
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 82
    Illegal SapI site found at 1191

Profile

Name: mchF
Base Pairs: 2097bp
Origin: Escherichia coli, synthesis
Properties: Microcin H47(MccH47) ABC transporter protein

Usage and Biology

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

Design and Construction

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


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.