| 鱼生长激素基因在大肠杆菌中表达水平的研究 |
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| 引用本文： | 白俊杰,马进,简清,李新辉,罗建仁,林文辉,张红军.鱼生长激素基因在大肠杆菌中表达水平的研究[J].中国水产科学,2000,7(1):31-34. |
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| 作者姓名： | 白俊杰 马进 简清 李新辉 罗建仁 林文辉 张红军 |
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| 作者单位： | 农业部,热带亚热亚热带鱼类选育与养殖重点开放实验室,中国水产科学研究院,珠江水产研究所,广东,广州,510380 |
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| 基金项目： | 农业部“九五”重点渔业科技项目 (渔 95 -B - 96 - 0 2- 0 2 - 0 4 )，中国水产科学研究院基金!资助项目 (95 - 0 8- 0 1) |
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| 摘 要： | 采用聚合酶链反应（ＰＣＲ）技术对鲤和鲑生长激素ｃＤＮＡ的５‘端和３’端进行定向改造。将改造后的５种基因分别克隆到大肠杆菌表达质粒ｐＢＶ２２０进行原核表达，以研究鱼生长激素基因在大肠杆菌中的表达水平。实验结果表明：（１）核糖核蛋白体结合位点（ＳＤ）与起始密码子ＡＵＧ之间的距离对鱼生长激素的表达水平有影响；（２）鲑鱼生长激素基因的终止密码子ＵＡＧ可能不会有效终止该基因在大肠杆菌中翻译的进行而造成部分通
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| 关 键 词： | 鱼生长激素 基因改造 表达水平 大肠杆菌 |
| 修稿时间： | 1999-04-23 |
Study on modification of fish GH cDNA and its expression in Escherichia coli |
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| BAI Jun-jie,MA Jin,JIAN Qing,LI Xin-hui,LOU Jian-ren,LIN Wen-hui,ZHANG Hong-jun.Study on modification of fish GH cDNA and its expression in Escherichia coli[J].Journal of Fishery Sciences of China,2000,7(1):31-34. |
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| Authors: | BAI Jun-jie MA Jin JIAN Qing LI Xin-hui LOU Jian-ren LIN Wen-hui ZHANG Hong-jun |
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| Abstract: | In order to study the expression level of fish growth hormone (fGH) in Escherichia coli, common carp and salmon GH cDNA's 5'ends and 3'ends were modified by using polymerase chain reaction (PCR) methods. Five of modified fGH cDNAs were coloned to E.coli expression vector pBV220. The experiments showed that the distance between SD sequence and initiator AUG is concerned with fGH expression level; salmon GH cDNA's stop codon UAG may not stop E. coli's translation effectively, but the strong stop condon UAA of E. coli added can avoid expression of super long proteins; although carp GH and salmon GH cDNA's 5'and 3'ends are modified with the same sequences,the expression levels of the 2 fish GH are quite different, showing that the number and distribution of rare codon in fGH genes may affect expression level of fGH in E.coli; the free energy of RNA 5'ends and local codon adaptation index also affect gene expression. |
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| Keywords: | fish growth hormone gene modification expression level Escherichia coli RNA secondary structure |
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