欢迎访问《中国农学通报》，
中国农学通报

中国农学通报 ›› 2009, Vol. 25 ›› Issue (16): 56-60.

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

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   

  • 收稿日期:2009-04-02 修回日期:2009-05-04 出版日期:2009-08-20 发布日期:2009-08-20

Construction and Expression of Prokaryotic Expression Vector for Sex-determining Gene Sry

Zhang Yanping, Zhang Yanquan, Guo Enmian, Zhu Baochang   

  • Received:2009-04-02 Revised:2009-05-04 Online:2009-08-20 Published:2009-08-20

PDF (PC)

888

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。本研究以小鼠为实验材料，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实验结果验证了蛋白表达的正确性。本实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实验基础。

关键词: 玉米秸秆, 玉米秸秆, 收刈时间, 有机物产量, 可消化有机物

Abstract: 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.