摘要
在毕赤酵母X33中表达人表皮生长因子并纯化,对表皮生长因子(EGF)进行密码子优化,构建pPICZa A-EGF真核表达质粒,转化X33感受态细胞;利用抗性筛选以及PCR鉴定阳性菌株。经过甲醇诱导和镍柱纯化,聚丙烯酰胺凝胶电泳检测蛋白分泌表达情况。成功构建了表达pPICZa A-EGF真核表达质粒,转入X33中获得阳性菌株,成功诱导蛋白分泌表达并纯化。在毕赤酵母X33中成功表达人表皮生长因子,纯化后纯度为80%,收率为5.8 mg/L。
Objective expression and purification of human epidermal growth factor in Pichia pastoris X33,methods egf was codon-optimized to construct pPICZa A-EGF eukaryotic expression plasmid and transformed into competent cells of X33. Positive strains were identified by resistance screening and PCR. After methanol induction and nickel column purification,SDS-PAGE was used to detection of protein secretion. The eukaryotic expression plasmid pPICZa A-EGF was successfully constructed and transformed into X33 to obtain a positive strain,which was successfully induced to express and purify protein. Human epidermal growth factor was successfully expressed in Pichia pastoris X33. The purity of purified protein was 80% with a yield of 5. 8 mg/L.
作者
高云鹏
赵雨
王新宇
白雪媛
王佳雯
GAO Yun-peng;ZHAO Yu;WANG Xin-yu;BAI Xue-yuan;WANG Jia-wen(Traditional Chinese Medicine and Biotechnology Research and Development Center,Changchun University of Chinese Medicine,Changchun 130117,China)
出处
《科学技术与工程》
北大核心
2018年第17期141-144,共4页
Science Technology and Engineering
基金
吉林省科技发展计划(20140622003JC)资助
关键词
表皮生长因子
毕赤酵母
蛋白表达
纯化
epidermal growtii factor
Pichic pastoris
protein expression
purification
