Construction of Recombinant Mice Factor Ⅶ-pPIC9K Vector and Expression in Pichia
- VernacularTitle:重组小鼠凝血因子Ⅶ-pPIC9K表达载体的构建及其在毕赤酵母中的表达
- Author:
Zhengrong YANG
;
Fei HE
;
Meng WANG
;
Yongping SHU
;
Tianming CHENG
- Publication Type:Journal Article
- Keywords:
coagulation factor Ⅶ;
tissue factor;
pichia pastoris;
site-directed mutagenesis
- From:
Chinese Journal of Cancer Biotherapy
1995;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct the yeast expressive vector of rmFⅦ, in which mFⅦ was mutated to inhibit coagulation without affecting the affinity for TF, and express it in Pichia pastoris. Methods:The full length cDNA encoding mFⅦ was amplified from a mouse liver by RT-PCR method, site-direct mutated and restriction enzyme digested as design. Cloning into pPIC9K, electroporation of Gs115, in vivo screen of multiple inserts by G418 resistance, BMGY/BMMY are used for induction and expression of rmFⅦ in pichia pastoris. These proteins were also screened for functional activity. Results: Three different rmFⅦ-pPIC9K yeast expression vectors and it's aim protein were obtained,two kinds of proteins were found to be functional active as design. Conclusion:rmFⅦ protein can be expressed in pichia pastoris and it might facilitate the development of tumor-target molecule, and novel anti-agiogenesis drug study.
