Cell

Part:BBa_K5246038

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


BL21(DE3) 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. BL21(DE3) is an E.coli expression strain suitable for the expression of non-toxic heterologous genes. The BL21(DE3) strain has a lambda DE3 prophage that allows for the expression of T7 RNA polymerase with IPTG. This strain is an E. coli B derivative that lacks the lon and OmpT proteases, which help preserve foreign proteins from degradation.

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)(KanR) ΔwecB

Experimental characterization

[edit]
Categories
Parameters
genotypeF- ompT hsdSB (rB-, mB-) gal dcm (DE3)(KanR) ΔwecB