The Research Of Prohibitin1 And B- Lymphocyte-like Immune Regulation In Lampetra Japonica

Prohibitin ( PHB) is a highly conserved protein and ubiquitously distributed in different cellular compartments of various organisms.Various roles have been proposed for prohibitin involving multibiological processes such as cell cycle regulation,cell proliferation,cell aging and cell apoptosis,etc. Due to its highly conserved structure in evolution and diverse functions, PHB has gradually caused widespread concern.The purpose of this study is to obtain the full-length cDNA encoding the Prohibitin protein. According to the information from cDNA library and the primary analysis of expressed sequence tags, In final, we also cloned a Prohibitin gene, which contained a 828bp open reading frame, and encoded 275 amino acids residues, from the leukopenia of lamprey, Lampetra japonica, which is one of the most primitive jawless vertebrates still living today.Then the cDNA was directionally cloned into the pET32a and transformed intoE. coli BL21, and expression was induced by IPTG. After the identification by SDS-PAGE, the recombinant proteins immunoblotting analysis to detecte the rcombinant protein . We take gene engineering mothod and the recombinant proteins were taken prokaryotic expression .We analysis the protein with tools of bioinformatics and check the specific expression in the different organization and Biological Activity of Prohibitin.We detected specific expression of the different organization using the method examination of reverse transcription PCR.The result expresses that this gene is an necessary important gene in most of organizations.The Prohibitin mRNA was also detected in lamprey gut, kidney, and leukocyte, but absent in lamprey buccal gland Prohibitin Speculated may be involved in a class B-lymphocyte-mediated signaling pathway.Design experiments,Using of RT-PCR method to detect different tissues before and after the PHA to stimulate the expression of the changes in Prohibitin1. The results show that PHA-stimulated leukocytes, heart, gill, their content in a relatively large increase This is also an indirect proof of results in a lamprey in this ancient vertebrate class B lymphocytes regulate the existence of the possible.