YI Jing1, HAO Xiaoª²Ke1, SU Mingª²Quan1, YUE Qiaoª²Hong1, LIU Jiaª²Yun1,ZHANG Rui2, LEI Yingª²Feng3

    1Department of Clinical Laboratories, Xijing Hospital, 2Departª²ment of Biochemistry £¦ Molecular Biology, 3Department of Microbiology, School of Basic Medicine, Fourth Military Medical University, Xi¬ðan 710033, China

    ¡¾Abstract¡¿ AIM: To express the human carboxypeptidase A1 (CPA1) active center protein in soluble form in pichia yeast. METHODS: Human CPA1 active center gene was amplified from pGEXª²CPA1 recombinant plasmid and subcloned into pPIC9K yeast expression vector. Recombinant vector pPIC9Kª²CPA1 active center was transformed into pichia SMD1168 by electroporation. Positive recombinant yeast strains were screened through nutrition defect and G418 resistance. The integrated multicopy recombinant yeast strains were induced for expression. RESULTS: The yeast recombinant expression vector pPIC9Kª²CPA1 active center was constructed and recombinant yeast strains which integrated over 16 recombinant vector copies were obtained. The human CPA1 active center protein was expressed in soluble form in pichia yeast after screened with G418. CONCLUSION: Human CPA1 active center protein has been expressed successfully in pichia yeast, which will be useful for further research on ADEPT of prospate. ......