The expression of rhCEMP1 with the eukaryotic expression vector pWX530 in yeast

Objective:To construct the recombinant eukaryotic expression vector containing the recombination human cementum protein 1(rhCEMP1) and to observe the expression of rhCEMP1 in yeast.Methods:RhCEMP1 cDNA was amplified by PCR,and correctly inserted into corresponding sites of intermediate vector pTeasy after restriction endonuclease digestion,and then inserted into corresponding sites of eukaryotic expression vertor pWX530 after another restriction endonuclease digestion.The recombinant vector pWX530-rhCEMP1 was confirmed to contain rhCEMP1 DNA sequence by agarose gel electrophoresis and DNA sequence analysis.The correct vector pWX530-rhCEMP1 was transfectd into yeast competent cells,and then the yeast was cultured after amino acid auxotroph screening.The expression of rhCEMP1 was determined by SDS-PAGE and enzyme-linked immunosorbent assay(ELISA),and the rhCEMP1 was purified by ion exchange chromatography.Results:The recombinant plasmid was successfully transferred to the yeast cells.The rhCEMP1 was successfully expressed by SDS-PAGE and ELISA analysis.Conclusions:The recombinant vector pWX530-rhCEMP1 was constructed successfully.The recombinant plasmid pWX530-rhCEMP1 could be transfected into yeast and expressed.