title: [Construction of vector of his-tagged cytoplasmic fragment of human toll like receptor 4 and its expression in E.coli].
abstract: OBJECTIVE
  To construction of vector of his-tagged cytoplasmic fragment of human Toll like receptor 4 (hTLR4) and its expression in E.coli.
  
  
  METHODS
  hTLR4 cytoplasmic cDNA codon domain was amplified by polymerase chain reaction (PCR) and cloned into pET-DsbA2.0 plasmid expressing His-DsbA fusion protein. After being identified by the assay of restrictional enzyme and sequencing, His-Dsb A fusion proteins were induced with isopropy-beta-D-thiogalactoside (IPTG) and further purified.
  
  
  RESULTS
  A fusion protein with molecular weight of 42 kd was obtained.
  
  
  CONCLUSIONS
  hTLR4 which was constructed and expressed successfully under nondenaturing conditions provides a tool for further studies.
authors.affiliation: Department of Pathophysiology and Key Laboratory of Shock and Microcirculation of People's Liberation Army, First Military Medical University, Guangzhou 510515, Guangdong, China.
authors.forename: Jing-hua
authors.initials: JH
authors.lastname: Liu
authors.forename: Xue-gang
authors.initials: XG
authors.lastname: Sun
authors.forename: Zhi-jie
authors.initials: ZJ
authors.lastname: Li
authors.forename: Qing-he
authors.initials: QH
authors.lastname: Qin
authors.forename: Xiao-wei
authors.initials: XW
authors.lastname: Gong
authors.forename: Peng
authors.initials: P
authors.lastname: Deng
authors.forename: Ya-wei
authors.initials: YW
authors.lastname: Liu
authors.forename: Jing-zhen
authors.initials: JZ
authors.lastname: Wang
authors.forename: Ke-sen
authors.initials: KS
authors.lastname: Zhao
authors.forename: Yong
authors.initials: Y
authors.lastname: Jiang
mesh_terms: D003001:Cloning, Molecular / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011956:Receptors, Cell Surface / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D051197:Toll-Like Receptor 4; D051193:Toll-Like Receptors
chemical_list: D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; C495345:TLR4 protein, human; D051197:Toll-Like Receptor 4; D051193:Toll-Like Receptors