2010•Zhongguo mianyixue zazhiRequires access
Expression of human ZP3 186S protein in Pichia pastoris X33
Hongjun Wang, Zhixin Li, Tian HongYan, Xu Ye, Xinwei Zhu, Lin DongLing, Zhao-hua Dou
Open publisher page 0 citations
Abstract
Objective:The study is aimed to express human ZP3 186S protein corresponding to 23-348 aa in Pichia pastoris,and to provide material in clinical diagnosis.Methods:The hZP3 cDNA encoding 23-348 aa was obtained by PCR and the expression plasmid pPICZα-hZP3 was constructed by inserting hZP3 cDNA.The expression plasmid pPICZα-hZP3 186S was generated by a mutation from arginine to serine in ZP3 and transformed into Pichia pastoris stain X33.The transformants were selected on media containing 2 mg/ml of Zeocin.Then hZP3 186S protein was induced for expression by methanol and analyzed by SDS-PAGE and Western blot.Results:The expression plasmid pPICZα-hZP3 186S was proved to be correct after sequencing,and hZP3 186S protein was efficiently expressed in Pichia pastoris.Conclusion:The hZP3 186S protein corresponding to 23-348 aa can be expressed in Pichia pastoris,and we will study its bioactivity to provide material for infertile diagnosis and immunocontraceptive vaccines.