摘要
目的研究干扰素HuIFNɑ-2b在酿酒酵母中分泌表达。方法采用PCR技术以IFNα-2b基因为模板定向克隆到pGAPZα-A表达载体中;转化到酿酒酵母菌进行表达,表达后的产物用SDS-PAGE蛋白质电泳鉴定,采用微量细胞病变抑制法进行测活。结果IFNα-2b基因克隆到表达质粒载体pGAPZα-A并成功地实现了在酿酒酵母中的表达,该目标蛋白不需复性,其活性可达到1.0×109IU/mg蛋白。结论IFNα-2b基因在酿酒酵母中成功地克隆表达,表达后的产物比大肠杆菌表达的HuIFNα-2b的活性高而且杂蛋白少,对规模化生产十分有利。
Objective Investigated the secretion expression of interferon HuIFN -2b in Saccharomyces(S.) cerevisiae. Method The target proteins were directionally cloned to pGAPZα-A expression vectors with IFNα-2b genes as templates by PCR technique; transferred to S. cerevisiae for expression, the expressed products were assayed via SDS-PAGE protein electrophoresis, the activity were determined with trace-level cell affection inhibition. Results IFNα-2b genes were cloned to the plasmid expression vectors pGAPZα-A and successfully expressed in S. cerevisiae; the target protein required no renaturation and its activity reached 1.0×109IU/mg protein. Conclusion IFNα-2b genes were successfully expressed in S. cerevisiae and the resulted activity was higher and impurity proteins were less than those expressed in E.colis., which will be of a great help in large scale production.
出处
《药品评价》
CAS
2004年第5期350-352,共3页
Drug Evaluation
关键词
干扰素Α-2B
酿酒酵母
基因表达
基因产物
蛋白质
interferon α-2b
Saccharomgces cerevisiac
gene expression
gene products, protein
