| 双须骨舌鱼卵黄蛋白原基因vtg的克隆、组织表达和原核表达分析 |
| |
| 引用本文： | 潘贤辉,刘奕,周康奇,胡隐昌,郑曙明,宋红梅,牟希东,杨叶欣.双须骨舌鱼卵黄蛋白原基因vtg的克隆、组织表达和原核表达分析[J].动物学杂志,2018,53(4):597-614. |
| |
| 作者姓名： | 潘贤辉 刘奕 周康奇 胡隐昌 郑曙明 宋红梅 牟希东 杨叶欣 |
| |
| 作者单位： | 西南大学淡水鱼类资源与生殖发育教育部重点实验室;农业部休闲渔业重点实验室广东省现代休闲渔业工程技术研究中心中国水产科学研究院珠江水产研究所 |
| |
| 基金项目： | 广州市科技计划项目（No. 201707010309），渔港建设和渔业产业发展专项（No. A201701A07），国家水产种质资源平台项目（No. 2018DKA30470），广东省自然科学基金项目（No. 2014A030310397） |
| |
| 摘 要： | 为探究双须骨舌鱼(Osteoglossum bicirrhosum)卵黄蛋白原vtg基因的结构和表达特征,本研究运用RACE技术克隆双须骨舌鱼vtg全长c DNA序列。序列分析发现,双须骨舌鱼vtg的c DNA全长为5 325 bp,开放阅读框(ORF)为5 121 bp,其5′和3′非编码区(UTR)分别为46 bp和159 bp,共编码1 706个氨基酸,预测蛋白质分子量约为186.79 ku。同源性分析发现,双须骨舌鱼与同科的美丽仆骨舌鱼(Scleropages formosus)相似度达到84.78%,与鲤形目、鲈形目、鲱形目、鲽形目鱼类的相似度均高于50%。系统进化树分析结果显示,本文获得的双须骨舌鱼vtg基因与骨舌鱼目聚为一类,与鳗鲡目硬骨鱼类亲缘关系较近,与双须骨舌鱼进化地位相符。荧光定量PCR检测结果表明,在雌雄鱼性腺和肝中vtg均有表达,且在雌鱼肝中的相对表达量显著高雄鱼(P0.05),卵巢和精巢表达量无显著差异(P0.05);在雌雄鱼鳃、脾、肾、肌肉、心、头肾、脑等7个组织中均极微量表达,且无显著差异(P0.05)。在Ⅱ、Ⅲ、Ⅳ期卵巢中vtg相对表达量依次增加,Ⅳ期显著高于Ⅱ和Ⅲ期(P0.05),也显著高于精巢Ⅳ期(P0.05);在雌鱼肝组织中呈先增后减趋势,且Ⅱ、Ⅲ、Ⅳ期之间无显著差异(P0.05),但每个时期都显著高于雄性(P0.05)。成功构建了原核表达载体p EASY-Blunt E2-vtg,并转入Transetta(DE3)表达感受态细胞中成功诱导出融合蛋白。Western-blotting检测显示,融合蛋白可被抗His标签特异性识别。综上表明,vtg表达水平高低与性别相关,并与性腺发育程度有关。此外,本研究结果也为后续深入研究卵黄蛋白原的生理功能打下基础。
|
| 关 键 词： | 双须骨舌鱼 卵黄蛋白原 克隆 组织表达 原核表达 |
| 收稿时间： | 2017/9/14 0:00:00 |
| 修稿时间： | 2018/6/22 0:00:00 |
Molecular Cloning, Expression Pattern and Prokaryotic Expression of Vitellogenin Gene (vtg) from Osteoglossum bicirrhosum |
| |
| Institution: | Key Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, China Southwest University, Chongqing 400715; Key Laboratory of Recreational Fisheries, Ministry of Agriculture, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380,Key Laboratory of Recreational Fisheries, Ministry of Agriculture, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380,Key Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, China Southwest University, Chongqing 400715; Key Laboratory of Recreational Fisheries, Ministry of Agriculture, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380,Key Laboratory of Recreational Fisheries, Ministry of Agriculture, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380,Key Laboratory of Recreational Fisheries, Ministry of Agriculture, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380,Key Laboratory of Recreational Fisheries, Ministry of Agriculture, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380,Key Laboratory of Recreational Fisheries, Ministry of Agriculture, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380 and Key Laboratory of Recreational Fisheries, Ministry of Agriculture, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380 |
| |
| Abstract: | |
| |
| Keywords: | Osteoglossum bicirrhosum Vitellogenin Cloning Tissue expression Prokaryotic expression |
| 本文献已被 CNKI 等数据库收录！ |
| 点击此处可从《动物学杂志》浏览原始摘要信息 |
| 点击此处可从《动物学杂志》下载免费的PDF全文 |
|
