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| 猪IL-18在毕赤酵母中分泌表达条件的优化 | |
| 引用本文： | 宋世斌,陈国华,王颖,王笑笑,宗瑞谦,景志忠.猪IL-18在毕赤酵母中分泌表达条件的优化[J].甘肃农业大学学报,2008,43(1):38-42. |
| 作者姓名： | 宋世斌 陈国华 王颖 王笑笑 宗瑞谦 景志忠 |
| 作者单位： | 1. 甘肃农业大学动物医学院,甘肃,兰州,730070;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 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 4. 甘肃农业大学动物医学院,甘肃,兰州,730070 |
| 基金项目： | 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. |
| 关 键 词： | 猪IL-18 毕赤酵母 摇瓶发酵 表达条件 |
| 文章编号： | 1003-4315（2008）01-0038-05 |
| 收稿时间： | 2007-03-15 |
| 修稿时间： | 2007年3月15日 |
Optimization of expression conditional optimization of porcine IL-18 expressed in Pichia Pastoris |
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| SONG Shi-bin,CHEN Guo-hua,WANG Ying,WANG Xiao-xiao,ZONG Rui-qian,JING Zhi-zhong.Optimization of expression conditional optimization of porcine IL-18 expressed in Pichia Pastoris[J].Journal of Gansu Agricultural University,2008,43(1):38-42. | |
| Authors: | SONG Shi-bin CHEN Guo-hua WANG Ying WANG Xiao-xiao ZONG Rui-qian JING Zhi-zhong |
| Abstract: | 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. |
| Keywords: | IL-18 Pichia pastoris shake-flask fermentation expression condition |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |