猪IL-18在毕赤酵母中分泌表达条件的优化  

Optimization of expression conditional optimization of porcine IL-18 expressed in Pichia Pastoris

作　　者：宋世斌[1,2] 陈国华[2] 王颖[3] 王笑笑[1,2] 宗瑞谦[1] 景志忠[2] 

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070 [2]中国农业科学院兰州兽医研究所,甘肃省动物寄生虫病重点实验室,家畜疫病病原生物学国家重点实验室,甘肃兰州730046 [3]新疆农业大学动物医学学院,新疆乌鲁木齐830052

出　　处：《甘肃农业大学学报》2008年第1期38-42,共5页Journal of Gansu Agricultural University

基　　金：兰州市科技攻关项目(05-1-47和06-2-23)资助.

摘　　要：采用PCR方法从pGEM-IL-18重组质粒中亚克隆出IL-18基因并完成了真核融合表达载体pPIC9K-IL-18的构建和IL-18在毕赤酵母中的表达,在此基础上从发酵时间、不同诱导型、pH值、诱导剂量和接种量等方面对重组基因工程菌的摇瓶发酵条件进行了优化,进一步探索了酵母菌表达猪IL-18的发酵工艺.结果表明,摇瓶发酵时,诱导最佳时间为72 h,甲醇最适浓度为20 g/L,最适pH范围为6.0-8.0,最适接种量1∶1.与单纯甲醇诱导相比,甘油/甲醇诱导后的表达量明显提高.The porcine IL-18 gene was amplified from recombinant plasmid pGEM-IL-18 by PCR, the positive expression vector, designated as pPIC9K-IL-18, was constructed and induced by methanol.Different culturing time, inducible pattern, pH and the amount of inducible reagents were compared, respectively. The results revealed that the optimal fermentation timing is was 72 h after induction with methanol,the optimal concentration of methanol was 20 g/L,the optimal pH range was 6.0-8.0 and the optimized inoculation amount was 1 ： 1. Compared with the induction only with methanol, combined use of glycerol and methanol could induce higher expression of the interest protein.

关 键 词：猪IL-18 毕赤酵母 摇瓶发酵 表达条件 

分 类 号：Q786[生物学—分子生物学]