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| 大菱鲆胰岛素样生长因子-I成熟肽的克隆、重组表达及活性分析 | |
| 引用本文： | 赵晓杰,陈松林,王娜,王贤丽,邢士超.大菱鲆胰岛素样生长因子-I成熟肽的克隆、重组表达及活性分析[J].水产学报,2010,34(1):1-7. |
| 作者姓名： | 赵晓杰 陈松林 王娜 王贤丽 邢士超 |
| 作者单位： | 1. 中国海洋大学海洋生命学院,山东,青岛,266003;中国水产科学研究院黄海水产研究所,农业部海洋渔业资源可持续利用重点开放实验室,山东,青岛,266071 2. 中国水产科学研究院黄海水产研究所,农业部海洋渔业资源可持续利用重点开放实验室,山东,青岛,266071;上海海洋大学,上海高校水产养殖学E-研究院,上海,201306 3. 中国水产科学研究院黄海水产研究所,农业部海洋渔业资源可持续利用重点开放实验室,山东,青岛,266071 |
| 基金项目： | 国家“八六三”高技术研究发展计划(2006AA10A402); 山东省泰山学者工程专项经费资助; 上海市教育委员会E-研究院建设项目(E03009) |
| 摘 要： | 通过RT-PCR方法从大菱鲆肝组织克隆了胰岛素样生长因子-I(IGF-I)成熟肽片段,分析表明,此成熟肽由70个氨基酸残基组成,含有3个链内二硫键。将扩增片段克隆到原核表达载体pGEX-4T-1上,实现了IGF-I成熟肽和GST蛋白在Escherichia coli BL21(DE3)plysS中的融合表达。融合蛋白分子量约为34ku,诱导4h时占菌体总蛋白的59%,主要以包涵体形式存在。Western-blotting免疫印迹表明,融合蛋白可以特异性地被anti-GST抗体识别。包涵体经6mol/L盐酸胍变性溶解及脉冲法稀释复性后,通过GSTrapFF亲和预装柱纯化,获得了电泳分析纯的融合蛋白。以细胞增殖实验检测蛋白生物活性,结果显示,纯化蛋白能促进大菱鲆肾脏细胞的增殖。 |
| 关 键 词： | 胰岛素样生长因子-I cDNA克隆 重组表达 包涵体 融合蛋白 |
| 收稿时间： | 2008/9/30 0:00:00 |
| 修稿时间： | 2009/1/17 0:00:00 |
Cloning,recombinant expression and purification of turbot(Scophthalmus maximus) mature IGF-I |
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| ZHAO Xiao-jie,CHEN Song-lin,WANG N,WANG Xian-li and XING Shi-chao.Cloning,recombinant expression and purification of turbot(Scophthalmus maximus) mature IGF-I[J].Journal of Fisheries of China,2010,34(1):1-7. | |
| Authors: | ZHAO Xiao-jie CHEN Song-lin WANG N WANG Xian-li and XING Shi-chao |
| Institution: | ZHAO Xiao-jie1,2,CHEN Song-lin2,3,WANG Na2,WANG Xian-li2,XING Shi-chao2 (1.College of Marine Life Sciences,Ocean University of China,Qingdao 266003,China,2.Key Lab for Sustainable Utilization of Marine Fisheries Resources,Ministry of Agriculture,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao 266071,3.The E-Institute of Shanghai Municipal Education Commission,Shanghai Ocean University,Shanghai 201306,China) |
| Abstract: | Insulin-like growth factor-I (IGF-I) is a conserved peptide expressed ubiquitously,which shows highly homologous diverse effects on development,growth,and metabolism. With RT-PCR,the fragment encoding the turbot (Scophthalmus maximus) mature IGF-I peptide was amplified. It was predicted that the mature peptide was composed of 70 amino acids including 6 cysteines which may form 3 disulfide bonds. The target fragment was then successfully subcloned into the express vector pGEX-4T-1 and was highly expressed in... |
| Keywords: | insulin-like growth factor-I cDNA cloning recombinant expression inclusion body recombinant protein |
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