Cell

Part:BBa_K5246039

Designed by: Edgaras Zaboras   Group: iGEM24_Vilnius-Lithuania   (2024-09-23)


Rosetta(DE3)pLysS ΔWecB

Introduction

Strain description

Usage and Biology

WecB is a UDP-N-acetylglucosamine 2-epimerase that catalyzes the first step in UDP-ManNAcUA(UDP-ManNAcUA) biosynthesis. UDP-ManNAcUA is one of the building blocks for the cell surface polysaccharide enterobacterial common antigen (ECA). WecB synthesizes one of the essential sugars for ECA, and deleting the wecA gene impairs the ECA synthesis pathway in E.coli, therefore lipopolysaccharides containing it are not produced. UDP-ManNAcUA is hypothetically one of the monosaccharides in polysaccharides produced by C. crescentus. We wanted to impair the synthesis of UDP-ManNAcUA to learn about the composition of our polysaccharide. Rosetta(DE3)pLysS strain is designed to overcome the limitations of E. coli's codon usage by providing additional tRNAs for certain codons found in eukaryotic proteins. This allows for better expression of these proteins. In Rosetta(DE3)pLysS, the rare tRNA genes and the T7 lysozyme gene are on the same plasmid.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Functional Parameters

genotypeF- ompT hsdSB(rB- mB-) gal dcm (DE3) pLysSRARE (CamR)(KanR) ΔwecB

Experimental characterization

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Categories
Parameters
genotypeF- ompT hsdSB(rB- mB-) gal dcm (DE3) pLysSRARE (CamR)(KanR) ΔwecB