Difference between revisions of "Part:BBa K3924047"

 
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<partinfo>BBa_K3924047 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3924047 SequenceAndFeatures</partinfo>
  
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==Profile==
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Name: mchEF<br/>
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Base Pairs: 3298bp<br/>
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Origin: Escherichia coli, merge mchE and mchF<br/>
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Properties: A polycistron can express both MchE and MchF. MchE and MchL transport mature MccH47 to the extracellular side.<br/>
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==Usage and Biology==
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The mchE and mchF are both from E.coli CA46. The mchE encodes MchE and the mchF encodes MchF. Both MchE and MchF transport mature MccH47 to the extracellular side.<sup>[1]</sup> The polycistron aim to make the engineering bacteria have the ability to transport MccH47.<br/>
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==Design and Construction==
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To construct the plasmid, we use endonuclease SpeI to cut RGP-mchE at 2bp after mchE open reading frame(ORF), then we used NEB Hifi DNA assembly to add RBS(ribosome binding site)-mchF between mchE and rrnB terminator.<br/>
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[[Image: T--Tsinghua--part_mchEF_construct.png|'''Figure 1: The design of mchEF''']]
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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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Revision as of 09:57, 21 October 2021


mchE-mchF polycistrons with Ptac lacO promoter

mchE-mchF polycistrons with Ptac lacO promoter

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1713
    Illegal PstI site found at 610
    Illegal PstI site found at 823
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1713
    Illegal PstI site found at 610
    Illegal PstI site found at 823
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1713
    Illegal BamHI site found at 237
    Illegal BamHI site found at 2380
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1713
    Illegal PstI site found at 610
    Illegal PstI site found at 823
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1713
    Illegal PstI site found at 610
    Illegal PstI site found at 823
    Illegal AgeI site found at 2761
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1347
    Illegal SapI site found at 2456

Profile

Name: mchEF
Base Pairs: 3298bp
Origin: Escherichia coli, merge mchE and mchF
Properties: A polycistron can express both MchE and MchF. MchE and MchL transport mature MccH47 to the extracellular side.

Usage and Biology

The mchE and mchF are both from E.coli CA46. The mchE encodes MchE and the mchF encodes MchF. Both MchE and MchF transport mature MccH47 to the extracellular side.[1] The polycistron aim to make the engineering bacteria have the ability to transport MccH47.

Design and Construction

To construct the plasmid, we use endonuclease SpeI to cut RGP-mchE at 2bp after mchE open reading frame(ORF), then we used NEB Hifi DNA assembly to add RBS(ribosome binding site)-mchF between mchE and rrnB terminator.
Figure 1: The design of mchEF


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.