重组DNA
大肠杆菌
生物
分子生物学
脂多糖
Toll样受体
TLR4型
受体
先天免疫系统
细胞生物学
化学
生物化学
基因
免疫学
作者
Isabelle Cognet,Amélie Benoit de Coignac,Giovanni Magistrelli,Pascale Jeannin,Jean‐Pierre Aubry,Karine Maisnier-Patin,Gersende Caron,Sylvie Chevalier,Frédéric Humbert,Thien Ngoc Nguyen,Alain Beck,Dominique Velin,Yves Delneste,Martine Malissard,Jean‐François Gauchat
标识
DOI:10.1016/s0022-1759(02)00506-9
摘要
Mutations in the Escherichia coli (E. coli) and Salmonella lpxM gene have been shown to result in strains which grow normally and which produce a non-myristoylated lipopolysaccharide (nmLPS) with strongly reduced endotoxicity. Using homologous recombination, we inactivated the lpxM gene in BL21 (DE3), a strain widely used for the production of recombinant proteins. This led to a derivative unaffected in its capacity to support the production of recombinant proteins. This new strain expresses non-myristoylated LPS that induces markedly less activation and maturation of monocyte-derived dendritic cells (DC), as assessed by nuclear translocation of nuclear factor kappa B (NF-kappaB), production of TNF-alpha and IL-8 or expression of CD86. Activation of the main signal transducing receptor for extracellular LPS, Toll like receptor (TLR) 4 in conjunction with the soluble accessory protein MD-2 was also markedly decreased. The modified BL21 strain represents a new application of lpxM inactivation for the expression of proteins to be tested on dendritic cells or other LPS sensitive cells/receptor complexes. It is likely to be useful for the identification of new proteins activating the innate immune response and to reducing the risk linked with low level of endotoxin contamination in therapeutic recombinant proteins.
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