Expression Of Several Yeast Proteins In Various Expression Systems And Antibodies Preparation With The Purified Proteins

With the deeply developed of molecular biology,the technologies,such as western blot,chromatin immunoprecipitation(chiP) and co-immunoprecipitation (co-IP) become the routine means for genetics study,the same requirement of those experiment assays is need to high potency antibody.Therefore,antibody preparation is the basic work for these studies.Recombination proteins are commonly used as antigen for antibody preparation. The aim of this paper is expression protein in various expression systems,and then preparation of our target protein.The main contents are illustrated as following:1,6 component proteins gene of yeast were successfully cloned into prokaryotic expression vector.Identify some strains which can express target protein induced by IPTG;2,According the expression form of target protein,choose the fit method for purification;3,We detected the serum getting from the immunized rabbit by western blot. The results display we've gain some antibodies of those proteins;4,The Epll gene can't expression in E.coli,was cloned into pEMBLyex4.We attempt to express the Epl1 gene in eukaryotic expression system,but the result is not ideal;And then express several forms of RPL36B getting from yeast;5,We construct the cell free protein synthesis system base on E.coli extract,for exploitation the platform of new form of protein expression system in our lab,as yet, no adequate protein has been detected.We attempet to express proteins in various expression systems,one hand is to product proteins as antigen for antibody preparation,the other hand is to develop the new platform of protein synthesis.It's useful for our further study.