[Construction of vector of his-tagged cytoplasmic fragment of human toll like receptor 4 and its expression in E.coli].
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.
Liu
Jing-hua
JH
Department of Pathophysiology and Key Laboratory of Shock and Microcirculation of People's Liberation Army, First Military Medical University, Guangzhou 510515, Guangdong, China.
Sun
Xue-gang
XG
Li
Zhi-jie
ZJ
Qin
Qing-he
QH
Gong
Xiao-wei
XW
Deng
Peng
P
Liu
Ya-wei
YW
Wang
Jing-zhen
JZ
Zhao
Ke-sen
KS
Jiang
Yong
Y
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
D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; C495345:TLR4 protein, human; D051197:Toll-Like Receptor 4; D051193:Toll-Like Receptors