Difference between revisions of "Part:BBa K5246037"
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===Sequence and Features===
 
===Sequence and Features===
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<partinfo>BBa_K5246035 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K5246037 SequenceAndFeatures</partinfo>
  
 
===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K5246035 parameters</partinfo>
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<partinfo>BBa_K5246037 parameters</partinfo>
  
 
===Experimental characterization===
 
===Experimental characterization===

Revision as of 14:10, 23 September 2024


HMS147(DE3) ΔWecA

Introduction

Strain description

Usage and Biology

WecA is Undecaprenyl-phosphate α-N-acetylglucosaminyl transferase initiating the biosynthesis of enterobacterial common antigen (ECA) and O-antigen PS by catalyzing the transfer of N-acetylglucosamine (GlcNAc)-1-phosphate onto undecaprenyl phosphate to form Und-P-P-GlcNAc. WecA is the first glycosyltransferase, and deleting the wecA gene impairs the ECA synthesis pathway in E.coli, therefore lipopolysaccharides containing it are not produced. We wanted to disable the ECA pathway to reduce the metabolic burden on the cell. We aimed to optimize polysaccharide production by creating a more efficient strain.HMS174(DE3) strain is an alternative expression strain to BL21(DE3). Retains the lon protease, which can degrade heterologous proteins. More efficient at expressing non-prokaryotic proteins with rare codons due to its different codon usage bias.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 301
    Illegal XbaI site found at 1523
    Illegal PstI site found at 958
    Illegal PstI site found at 1545
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 958
    Illegal PstI site found at 1545
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1171
    Illegal BamHI site found at 1557
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 301
    Illegal XbaI site found at 1523
    Illegal PstI site found at 958
    Illegal PstI site found at 1545
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 301
    Illegal XbaI site found at 1523
    Illegal PstI site found at 958
    Illegal PstI site found at 1545
    Illegal NgoMIV site found at 507
  • 1000
    COMPATIBLE WITH RFC[1000]

Functional Parameters

genotypeF- recA1 hsdR(rK12- mK12+) (DE3) (Rif R)(KanR) ΔwecA

Experimental characterization