<正>Objective: To explore high yield secretory expression of rccombinant mouse coagulation factor VII （rmFVII ） protein in Pichia pastoris （P. pastoris）. Methods: The fragment of mFVII cDNA was amplified by PCR from a pcDNA3 mFVII plasmid. Then the cDNA fragment was suhclonecl intoα-factor secretion signal open reading frame of pPIC9K secretory expression vector. The mutagenesis of mFVII was performed by Site-Direct Mutation and then verified by DNA sequencing. The yeast expression vector of rmF VII. named as pPIC9K-rmF VII . was linearized with Sac I and transferred into GS115 strains（his Mut+ ） by electroporation. The rccombinanls were identified by direct PC’R and selection on MM and MD plates. rmFVII was expressed in recombinant strains （his Mut ） for 4 d. The expression level and activation of rmFVII in the BMMY medium were detected by SDS PAGE and Western blot respectively. Results: pPIC9K-rmF VII was constructed and transferred to GS115 strains successfully. 48 hour post induction by methanul rmFII protein was secreted into the culture supernatant. The molecular weight of the expressed produets was shown to he about 46 kD by SDS PAGE analysis. Western blot showed that the expressed rmF VII exhibited specificity and antigenicity. Conclusion: Since mF VII is considered as a tumor-targeting molecule

this study may provide a basis for further ami tumor strategy on rmFVII.