Human carboxypeptidase A1 active site of soluble expression in Pichia pastoris.docVIP

 PAGE \* MERGEFORMAT 12 Human carboxypeptidase A1 active site of soluble expression in Pichia pastoris Authors: Yi Jing, Hao Ke, Su rights, YUE Qiao-hong, LIU Jia-yun, Zhang Rui, Lei Ying-feng [Keywords:] human carboxypeptidase [Abstract] 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 pGEXCPA1 recombinant plasmid and subcloned into pPIC9K yeast expression vector. Recombinant vector pPIC9KCPA1 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 pPIC9KCPA1 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. [Keywords] carboxypeptidase A; active center; pichia; gene expression [Abstract] Objective: The Pichia pastoris expression system expression of human carboxypeptidase A1 (carboxypeptidase A1, CPA1) protein active site. Methods: From pGEXCPA1 recombinant active site was amplified by CPA1 gene, was subcloned into the yeast expression vector pPIC9K, recombinant yeast expression vector pPIC9KCPA1active center transformed into Pichia pastoris electric SMD1168, and screening for nutritional deficiencies and the G418 resistance screening for high-copy integration of the recombinant yeast strain expressing inducible expression. Results: Construction of fusion recombinant yeast expression vectors are pPIC9KCPA1active center, through the G418 concentration gradient filter to b