| 类蚕丝丝素结晶区蛋白表达载体的构建与表达 |
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| 引用本文： | 王建南,田保中,左保齐,李明忠,吴徵宇,黄君霆,陆长德,白伦. 类蚕丝丝素结晶区蛋白表达载体的构建与表达[J]. 蚕业科学, 2006, 32(1): 36-41 |
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| 作者姓名： | 王建南 田保中 左保齐 李明忠 吴徵宇 黄君霆 陆长德 白伦 |
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| 作者单位： | 苏州大学材料工程学院,苏州,215021;苏州大学生命科学学院,苏州,215123;苏州大学材料工程学院,苏州,215021;中国农业科学院蚕业研究所,镇江,212018;中国科学院上海生命科学学院生物化学与细胞生物学研究所,上海,200031 |
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| 基金项目： | 中国科学院资助项目 , 江苏省苏州市农业科技基金 |
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| 摘 要： | 为探讨蚕丝结构与性能的关系,设计了几种类似蚕丝丝素结晶区高度重复序列结构(GAGAGX)n的基因序列,并构建了大肠杆菌表达载体,在BL-21菌株中成功诱导表达了4种约由110个氨基酸组成的小分子蛋白。对表达产物进行W estern印迹分析和ELISA检测的结果显示,(GAGAGA)n、(GAGAGY)n、(GAGAGV)n3种蛋白都有小于14.4 kD的大小相同的条带,而(GAGAGS)n有一条稍大于20.1 kD的条带。以超声波破菌的缓冲液作空白对照,4种蛋白的ELISA的结果均比对照的BL-21(无质粒)高,显示为阳性。
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| 关 键 词： | 仿丝素结晶区 载体构建 大肠杆菌表达 |
| 文章编号： | 0257-4799(2006)01-0036-06 |
| 收稿时间： | 2005-09-12 |
| 修稿时间： | 2005-09-12 |
Construction of Expression Plasmid Encoding Fibroin Crystalline Domain Like Protein of Silkworm and its Expression in E.coli |
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| WANG Jian-Nan,TIAN Bao-Zhong,ZUO Bao-Qi,LI Min-Zhong,WU Zheng-Yu,HUANG Jun-Ting,LU Chang-De,BAI Lun. Construction of Expression Plasmid Encoding Fibroin Crystalline Domain Like Protein of Silkworm and its Expression in E.coli[J]. Acta Sericologica Sinica, 2006, 32(1): 36-41 |
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| Authors: | WANG Jian-Nan TIAN Bao-Zhong ZUO Bao-Qi LI Min-Zhong WU Zheng-Yu HUANG Jun-Ting LU Chang-De BAI Lun |
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| Affiliation: | 1.College of Material Engineering, Soochow University, Suzhou Jiangsu 215021, China; 2 .College of Life Science, Soochow University, Suzhou Jiangsu 215123, China; 3 .The Sericultural Research Institute, Chinese Academy of Agricultural Science, Zhenjiang Jiangsu 212018, China; 4.Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China |
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| Abstract: | We have designed four gene fragments encoding silkworm fibroin crystalline domain like proteins,(GAGAGX)_(n),which are highly repetitive in silk.These fragments were cloned into pCDNA3 expression vectors and successfully expressed in E.coli BL-21 under inducement of IPTG.The vectors were characterized by enzyme digestion and gene sequencing.Western blot analysis showed that three expressed proteins have molecular masses about 14.4 kD,another is a bit larger than 20.1 kD.ELISA analysis showed that all these four proteins extracted by ultrasonic were positive. |
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| Keywords: | Fibroin crystalline domain like protein Vector construction Gene expression in E.coli |
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