Prokaryotic Expression And Preliminary Function Research Of Molecular Chaperone JIV90 Gene

The molecular chaperone (molecular chaperones) are a group of widespreads from bacteria to human proteins, non-covalent reconciliation with the nascent chain folding of proteins chain binding, and help them fold and transport, usually not involved in the physiological functions of target proteins. Lackner et al found that the molecular chaperone Jiv90 of host cell interacts with CSFV NS2 proteins that has its own protease actiity, then NS2-3 protein complex cut into NS2 and NS3 protein monomers. Although there are some reports about Jiv90 genes but only limited interaction with the virus and viruses titer change. there is little research reports about Jiv90 protein functions. This paper reveals Jiv90 protein function as the goal, engaging in Jiv90 protein degradation characteristics, the distribution of Jiv90 gene on cell and organization of swine and other researchs.From the experiments, the following results were obtained:(1) The Jiv90 gene of was amplified by PCR with specific primers, and cloned into pET-32a(+) after cleavage by corresponding enzymes. The recombinant plasmid was named pET-32a-Jiv90. SDS-PAGE and western-blot showed an expected protein of 45 ku in size was successfully expressed in E. coli BL21(DE3) induced by IPTG.(2) The Jiv90 protein was expressed as fused to the green fluorescent protein (GFP), Western blot analysis showed that the molecular weight of Jiv90 was approximately 52 ku, this protein was a protein for its degradation through the proteasomal degradation pathway which was inhibited by proteasomal inhibitor MG132.(3) We obtained successfully distribution of the Jiv90 genes in cells and tissues of porcine by Real-time PCR. The distribution of Jiv90 gene in porcine vascular endothelial and pig kidney cells is higher than other cells and the distribution of Jiv90 gene in lung is higher than other tissues.