Difference between revisions of "Part:BBa K3924022"

 
 
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<partinfo>BBa_K3924022 short</partinfo>
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3924022 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3924022 SequenceAndFeatures</partinfo>
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==Profile==
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Name: mchD<br/>
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Base Pairs: 453bp<br/>
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Origin: Escherichia coli, synthesis<br/>
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Properties: Microcin H47(MccH47) acyltransferase<br/>
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==Usage and Biology==
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mchD was obtained from E. coli strain CA46. It encodes MchD which modifies the precursor of MccH47.<sup>[1]</sup><br/>
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==Design and Construction==
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We introduce mchD gene from E.coli CA46 into E.coli Nissle 1917. The sequence was obtained from E. coli CA46 microcin operon(GenBank: AJ515251.1). The mchD gene is added into lac operator(on RGP plasmid) so that the mchD 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_mchD_construct.png|center|600px|thumb|'''Figure 1: The design of mchD''']]
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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 07:24, 21 October 2021


mchD. MchD modify the precursor of MccH47.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 256
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 256
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 256
    Illegal BamHI site found at 249
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 256
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 256
  • 1000
    COMPATIBLE WITH RFC[1000]

Profile

Name: mchD
Base Pairs: 453bp
Origin: Escherichia coli, synthesis
Properties: Microcin H47(MccH47) acyltransferase

Usage and Biology

mchD was obtained from E. coli strain CA46. It encodes MchD which modifies the precursor of MccH47.[1]

Design and Construction

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


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.