|
|||||
|
|
| 猪IL-18在毕赤酵母中的表达及活性分析 | |
| 引用本文： | 宋世斌,景志忠,陈国华,王笑笑,宗瑞谦.猪IL-18在毕赤酵母中的表达及活性分析[J].生物工程学报,2007,23(5):818-823. |
| 作者姓名： | 宋世斌 景志忠 陈国华 王笑笑 宗瑞谦 |
| 作者单位： | 1. 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,兰州,730046;甘肃农业大学动物医学院,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,兰州,730046 3. 甘肃农业大学动物医学院,兰州,730070 |
| 基金项目： | 国家高技术研究发展计划(863计划);甘肃省兰州市科技攻关项目 |
| 摘 要： | 采用PCR方法从pGEM-IL-18重组质粒中扩增出IL-18基因并构建真核融合表达载体pPIC9K-IL-18,电激法转化入毕赤酵母GS115,采用G418抗性梯度法筛选得到多拷贝重组菌株,甲醇诱导表达,应用SDS-PAGE分析重组蛋白的表达情况,并将表达蛋白用凝胶层析柱纯化后,用MTT法检测其生物学活性。实验结果表明重组的GS115酵母菌株可表达分泌pIL-18,其表达在72h时达高峰,分泌量可达160mg/L,纯化的重组pIL-18蛋白具有显著的促进淋巴细胞增殖的活性,说明本试验已在毕赤酵母中在国内首次成功表达了具有生物学活性的pIL-18。 |
| 关 键 词： | 毕赤酵母 基因表达 纯化 生物活性 |
| 文章编号： | 1000-3061（2007）05-0818-06 |
| 修稿时间： | 2007-01-17 |
Expression and Analysis of Recombinant pIL-18 in Pichia pastoris |
|
| SONG Shi-Bin,JING Zhi-Zhong,CHEN Guo-Hu,WANG Xiao-Xiao and ZONG Rui-Qian.Expression and Analysis of Recombinant pIL-18 in Pichia pastoris[J].Chinese Journal of Biotechnology,2007,23(5):818-823. | |
| Authors: | SONG Shi-Bin JING Zhi-Zhong CHEN Guo-Hu WANG Xiao-Xiao and ZONG Rui-Qian |
| Institution: | 1 Key Laboratory of Veterinary Porasitology of Gansu Province/State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute,Chinese Acadamy of Agricultural Science,Lanzhou 730046,China2 College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China |
| Abstract: | The porcine IL-18 gene was amplified from recombinant plasmid pGEM-IL-18 by PCR,then the pPIC9K-IL-18 of fusion expression vector was constructed by inserting IL-18 fragment,and was transformed to GS115 by electroporation,multi-copy recombinant strains were screened by G418.The expression of recombinant fusion protein was induced by methanol,SDS-PAGE was used to analyze expression product,fusion protein was purified by Sephadex G-100 column,bioactivity of IL-18 was measured by MTT assays.Experiment results show fusion protein of pIL-18 secreted by GS115,expression reaches the secretion peak of 160mg/L at 72h.We have expressed and purified successfully the recombinant pIL-18 with obvious biological activity in Pichia pastoris. |
| Keywords: | pIL-18 |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |
| 点击此处可从《生物工程学报》浏览原始摘要信息 | |
| 点击此处可从《生物工程学报》下载免费的PDF全文 | |