论文部分内容阅读
目的建立毕赤酵母表达PSA系统,获高活性人前列腺特异性抗原,为临床相关疾病的监测,诊断和治疗提供关键材料。方法构建表达载体pPICZα-mPSA质粒,采用电转法,将pPICZα-mPSA质粒转染毕赤酵母X33细胞,Zeocin筛选转染细胞,甲醇诱导X33细胞表达rPSA,ELASA试剂盒筛选高表达rPSA的细胞株。结果构建了表达载体pPICZα-mPSA,获得了高表达rPSA的酵母X33细胞株,表达量为1.2mg/L。结论应用酵母表达体系成功表达人前列腺特异性抗原。
Objective To establish a Pichia pastoris-expressing PSA system and to obtain highly active human prostate-specific antigens to provide key materials for the monitoring, diagnosis and treatment of clinically relevant diseases. Methods Plasmid pPICZα-mPSA was constructed by electroporation. The pPICZα-mPSA plasmid was transfected into Pichia pastoris X33 cells. Zeocin was used to screen the transfected cells. Methanol-induced XPS cells expressed rPSA, and ELASA kit was used to screen rPSA-expressing cell lines. Results The expression vector pPICZα-mPSA was constructed and the yeast X33 cell line highly expressing rPSA was obtained. The expression level was 1.2 mg / L. Conclusion The human prostate-specific antigen was successfully expressed in yeast expression system.