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| 藏猪兴奋性氨基酸转运载体EAAC1基因的克隆与原核表达 | |
| 引用本文： | 傅德智,孔祥峰,杨焕胜,褚武英,李铁军,印遇龙. 藏猪兴奋性氨基酸转运载体EAAC1基因的克隆与原核表达[J]. 农业现代化研究, 2011, 32(6): 756-760 |
| 作者姓名： | 傅德智 孔祥峰 杨焕胜 褚武英 李铁军 印遇龙 |
| 作者单位： | 1. 中国科学院亚热带农业生态研究所,亚热带农业生态过程重点实验室/湖南省畜禽健康养殖工程技术研究中心,湖南长沙410125;中国科学院研究生院,北京100049 2. 中国科学院亚热带农业生态研究所,亚热带农业生态过程重点实验室/湖南省畜禽健康养殖工程技术研究中心,湖南长沙410125 3. 中国科学院亚热带农业生态研究所,亚热带农业生态过程重点实验室/湖南省畜禽健康养殖工程技术研究中心,湖南长沙410125;长沙大学生物工程与环境科学系,湖南长沙410003 |
| 基金项目： | 国家自然科学基金，中国科学院知识创新工程重要方向项目 |
| 摘 要： | 兴奋性氨基酸转运载体EAAC1是肠道内谷氨酸的主要载体.本研究根据人的基因编码区设计一对特异性引物,以藏猪空肠总RNA为模板,通过RT-PCR获得一长约1600 bp的cDNA片段,T/A克隆后测序,并进行序列分析；将该基因片段连接到原核表达载体pET-32a(+)中构建融合表达质粒,转化到E.coli BL21(DE3)中进行表达.测序结果显示,获得的cDNA全长为1575 bp,编码524个氨基酸；EAAC1分子量为57 kD,等电点为5.34,GenBank登录号为GQ375513;该蛋白质具有3个N-糖基化位点、8个蛋白激酶C磷酸化位点和1个cAMP/cGMP依赖性蛋白激酶磷酸化位点,含有7个跨膜结构域和一个信号肽序列；SDS-PAGE电泳检测结果表明,表达蛋白质与预期蛋白质大小一致.上述结果为进一步研究EAAC1的功能及其调控奠定了基础. |
| 关 键 词： | 藏猪 兴奋性氨基酸转运载体 序列分析 原核表达 |
Gene Cloning and Prokaryotic Expressing of Excitatory Amino Acid Carrier 1 in Tibetan Pig |
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| FU De-zhi,KONG Xiang-feng,YANG Huan-sheng,CHU Wu-ying,LI Tie-jun and YIN Yu-long. Gene Cloning and Prokaryotic Expressing of Excitatory Amino Acid Carrier 1 in Tibetan Pig[J]. Research of Agricultural Modernization, 2011, 32(6): 756-760 | |
| Authors: | FU De-zhi KONG Xiang-feng YANG Huan-sheng CHU Wu-ying LI Tie-jun YIN Yu-long |
| Affiliation: | Key Laboratory for Agro-ecological Processes in Subtropical Region/Hunan Engineering and Research Center of Animal and Poultry Science,Institute of Subtropical Agriculture,the Chinese Academy of Sciences,Changsha,Hunan 410125,China;Graduate University of Chinese Academy of Sciences,Beijing 100049,China;Key Laboratory for Agro-ecological Processes in Subtropical Region/Hunan Engineering and Research Center of Animal and Poultry Science,Institute of Subtropical Agriculture,the Chinese Academy of Sciences,Changsha,Hunan 410125,China;Key Laboratory for Agro-ecological Processes in Subtropical Region/Hunan Engineering and Research Center of Animal and Poultry Science,Institute of Subtropical Agriculture,the Chinese Academy of Sciences,Changsha,Hunan 410125,China;Graduate University of Chinese Academy of Sciences,Beijing 100049,China;Key Laboratory for Agro-ecological Processes in Subtropical Region/Hunan Engineering and Research Center of Animal and Poultry Science,Institute of Subtropical Agriculture,the Chinese Academy of Sciences,Changsha,Hunan 410125,China;Department of Biotechnology and Environmental Science,Changsha University,Changsha,Hunan 410003,China;Key Laboratory for Agro-ecological Processes in Subtropical Region/Hunan Engineering and Research Center of Animal and Poultry Science,Institute of Subtropical Agriculture,the Chinese Academy of Sciences,Changsha,Hunan 410125,China;Key Laboratory for Agro-ecological Processes in Subtropical Region/Hunan Engineering and Research Center of Animal and Poultry Science,Institute of Subtropical Agriculture,the Chinese Academy of Sciences,Changsha,Hunan 410125,China |
| Abstract: | |
| Keywords: | Tibetan pig excitatory amino acid carrier 1 sequence analysis prokaryotic expression |
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