| 沼泽型水牛卵泡抑素基因的真核表达研究 |
| |
| 引用本文： | 邓继贤,杨秀荣,韦英明,蒋和生.沼泽型水牛卵泡抑素基因的真核表达研究[J].中国畜牧兽医,2011,38(9):57-60. |
| |
| 作者姓名： | 邓继贤 杨秀荣 韦英明 蒋和生 |
| |
| 作者单位： | 1. 广西科学院中国东盟(广西)生物技术合作研发中心,广西南宁 530003;2. 广西大学动物科技学院,广西南宁 530005 |
| |
| 基金项目： | 广西自然科学基金(桂科自0640014); 科技创新能力与条件建设(桂科能0992028-9) |
| |
| 摘 要： | 试验通过候选基因法,选定卵泡抑素(follistatin,FS)作为影响水牛繁殖性能的主要候选基因,通过基因工程的方法,用XhoⅠ、SacⅡ双酶切广西大学动物遗传育种与繁殖实验室克隆的添加有ACC的Kozaka 序列的pMD-bFS质粒和pEGFP-N1,构建pEGFP-bFS真核表达质粒,将其与阳离子脂质体混匀后,分别转染体外培养的水牛胎儿成纤维(BFF)细胞和仓鼠肾(BHK)细胞系,经过48 h培养后,在相差荧光显微镜下观察绿色荧光蛋白的表达水平,用RT-PCR和Western blotting方法对转入质粒表达进行定性鉴定。结果显示,本研究成功构建了添加有ACC的Kozaka 序列的pEGFP-bFS真核表达质粒,该重组质粒在BFF和HBK两种细胞均表达,但在HBK细胞系的表达量稍高。本研究结果将为下一步制备具有生物活性的bFS工程疫苗,研究bFS对水牛繁殖性能的影响提供参考。
|
| 关 键 词： | 沼泽型水牛 卵泡抑素 真核表达 繁殖性能 |
| 收稿时间： | 2011-03-02 |
Eukaryotic Expression of Swamp Buffalo Follistatin |
| |
| DENG Ji-xian,YANG Xiu-rong,WEI Ying-ming,JIANG He-sheng.Eukaryotic Expression of Swamp Buffalo Follistatin[J].China Animal Husbandry & Veterinary Medicine,2011,38(9):57-60. |
| |
| Authors: | DENG Ji-xian YANG Xiu-rong WEI Ying-ming JIANG He-sheng |
| |
| Institution: | 1. China-ASEAN (Guangxi) Biotechnology Cooperative Center, Guangxi Academy of Sciences, Nanning 530003, China;2. College of Animal Science and Technology, Guangxi University, Nanning 530005, China |
| |
| Abstract: | In this study,follistatin gene was chosen as the candidate gene affecting the reproductive ability in buffalo. Recombinant plasmid pEGFP-bFS was conducted and expressed in both buffalo fetal fibroblast (BFF) and BHK cells. pMD-bFS and pEGFP-N1 were digested with XhoⅠ,Sac Ⅱ,then eukaryotic expression vector-pEGFP-bFS was constructed. Buffalo fetal fibroblast and BHK cells were cultured with pEGFP-bFS intermixing LipofectamineTM 2000 in carbon dioxide incubator with 37 ℃,5% CO2,100% humidity. After 24 h,expression levels of recombinant vector were verified through observing directly under phase contrast fluorescence microscope,RT-PCR and Western blotting. The results indicated that bFST expressed in both buffalo fibroblast and BHK cells. This study was a very significance to produce bioactive bFS engineered plasmid and research effects of bFS on reproductive ability. |
| |
| Keywords: | swamp buffalo follistatin eukaryotic expression reproductive ability |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
| 点击此处可从《中国畜牧兽医》浏览原始摘要信息 |
| 点击此处可从《中国畜牧兽医》下载免费的PDF全文 |
|
