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| 大口黑鲈肌肉生长抑制素前肽真核表达载体的构建及其在肌肉组织中的表达 | |
| 引用本文： | 张宁宁,白俊杰,何小平,郭玉函,叶星,李胜杰,杜晓东.大口黑鲈肌肉生长抑制素前肽真核表达载体的构建及其在肌肉组织中的表达[J].广东海洋大学学报,2009,29(1). |
| 作者姓名： | 张宁宁 白俊杰 何小平 郭玉函 叶星 李胜杰 杜晓东 |
| 作者单位： | 1. 广东海洋大学水产学院,广东,湛江,524088;中国水产科学研究院珠江水产研究所中国水产科学研究院热带亚热带鱼类选育与养殖重点开放实验室,广东,广州,510380 2. 中国水产科学研究院珠江水产研究所中国水产科学研究院热带亚热带鱼类选育与养殖重点开放实验室,广东,广州,510380 3. 广东海洋大学水产学院,广东,湛江,524088 |
| 基金项目： | 国家科技支撑项目，国家科技基础条件平台工作项目，广东省自然科学基金，中国水产科学研究院水产种质资源与养殖技术重点开放实验窒开放基金 |
| 摘 要： | 将大口黑鲈(Micropterus salmoide)肌肉生长抑制素(Myostatin,MSTN)前肽(MSTN-Pro)的cDNA定向克隆到真核表达载体pcDNA3.1(-)/mycHisB中,双酶切检测和测序鉴定证实,插入pcDNA3.1(-)/mycHisB载体中的片段为目的基因的核苷酸序列,MSTN基因前肽cDNA为正向插入,且重组质粒无错配或插入移位等突变。采用肌肉注射法将重组表达质粒注入大口黑鲈背部肌肉组织,在注射后第2天经RT-PCR检测到MSTN前肽基因mRNA的表达,第6天经免疫组化学检测到MSTN前肽蛋白的表达,第8天蛋白表达强度增强,对照组始终未检测到MSTN前肽基因的表达。 |
| 关 键 词： | 大口黑鲈 肌肉生长抑制素(MSTN) 真核表达 表达载体 肌肉注射 免疫组织化学 |
Construction of Eukaryotic Expression Vectors of Myostatin Propeptide Large-Mouth Bass (Micropterus salmoides) and Its Expression in Dorsal Muscle |
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| ZHANG Ning-ning,BAI Jun-jie,HE Xiao-ping,GUO Yu-han,LI Sheng-jie,YE Xing,DU Xiao-dong.Construction of Eukaryotic Expression Vectors of Myostatin Propeptide Large-Mouth Bass (Micropterus salmoides) and Its Expression in Dorsal Muscle[J].Journal of Zhanjiang Ocean University,2009,29(1). | |
| Authors: | ZHANG Ning-ning BAI Jun-jie HE Xiao-ping GUO Yu-han LI Sheng-jie YE Xing DU Xiao-dong |
| Abstract: | The largemouth bass(Micropterus salmoides) MSTN N-terminal propeptide(MSTN-Pro) cDNA was cloned and inserted into the expression vector of pcDNA3.1(-)/myc-HisB.The recombiant plasmid was identified by restriction enzymes and sequencing.The results suggested that the nucleotide sequence isolated from the recombinant pcDNA3.1(-)/mycHisB-MSTN-Pro was the same as expected.Then the recombinant plasmid pcDNA3.1(-)/mycHisB-MSTN-Pro was injected into the dorsal muscle of largemouth bass.On the 2nd day postinjection into the skeletal muscle,RT-PCR showed that the exogenous gene was transcribed.On the 6th day the protein could be detected at the injection site by immunohistochemistry and the protein expression became stronger on the 8th day than that on the 6th day.The MSTN propeptide was not expressed in the corresponding control.The MSTN propeptide gene eukaryotic expression plasmid pcDNA3.1(-)/mycHisB-MSTN-Pro is constructed successfully,and is expressed in vivo. |
| Keywords: | largemouth bass(Micropterus salmoides) Myostatin(MSTN) eukaryotic expression expression vector muscle injection immunohistochemistry |
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