| 性别决定基因Sry原核表达载体的构建和蛋白检测 | |
| 张艳萍 ; 张衍泉 ; 郭恩棉 ; 朱宝长 ; Zhang Yanping ; Zhang Yanquan ; Guo Enmian ; Zhu Baochang | |
| 2010-07-19 ; 2010-07-19 | |
| 关键词 | 性别决定 Sry 原核表达载体 蛋白表达 sex determination, Sry, prokaryotic expression vector, protein expression S814.7 |
| 其他题名 | Construction and Expression of Prokaryotic Expression Vector for Sex-determining Gene Sry |
| 中文摘要 | 动物的性别在畜牧业是一项很重要的经济性状,而哺乳动物的性别决定受睾丸决定基因Sry的启动,获得SRY蛋白并探索它与其他相关因子之间的相互关系对于深入了解该因子的作用机理和寻求高效经济的性别控制方法尤为重要。以小鼠为试验材料,通过PCR、质粒重组、酶切和测序等方法构建小鼠Sry的原核表达载体,使用IPTG诱导蛋白表达、SDS-PAGE电泳和western blotting等方法检测重组表达载体的原核表达。PCR、酶切和测序结果一致显示成功地构建了原核表达载体pET-21b-Sry;使用anti-Sry和anti-His特异性抗体进行的western blotting试验结果验证了蛋白表达的正确性。该试验不仅实现了哺乳动物性别决定因子的Sry原核表达,而其还获得了具有生物活性的SRY蛋白,对于深入研究Sry及其他相关基因的相互作用奠定了试验基础。; Sex of breeded animal is a very important character for stockbreeding, while for mammal, the sex determination is started by gene Sry. So, to obtain SRY protein and study it’s interaction with other correlative genes plays an important role in making up efficient and economic technique for sex control of breeded animal.In this experiment, we used PCR、plasmid recombination、restriction digest and sequencing to construct pET-21b-Sry for sex determining gene of mice – Sry and detect it’s validity. While, we adopt protein expression induced by IPTG、 SDS-PAGE and western-blotting to achieve and detect SRY protein from prokaryotic expression. All the results of PCR、plasmid recombination、restriction digest and sequencing displayed Sry fragement from mice and prokaryotic expression vector pET-21b-Sry were abteined succesfully. Then, the validity of Sry-His protein in BL21(DE3) was proved with western blotting using anti-Sry and anti-His. In conclusion, Prokaryotic Expression for Sry-His was carried out in this experiment. |
| 语种 | 中文 ; 中文 |
| 内容类型 | 期刊论文 |
| 源URL | [http://hdl.handle.net/123456789/77450]  |
| 专题 | 清华大学 |
| 推荐引用方式 GB/T 7714 | 张艳萍,张衍泉,郭恩棉,等. 性别决定基因Sry原核表达载体的构建和蛋白检测[J],2010, 2010. |
| APA | 张艳萍.,张衍泉.,郭恩棉.,朱宝长.,Zhang Yanping.,...&Zhu Baochang.(2010).性别决定基因Sry原核表达载体的构建和蛋白检测.. |
| MLA | 张艳萍,et al."性别决定基因Sry原核表达载体的构建和蛋白检测".(2010). |
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