CAJ | 학술논문

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Sty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Sty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-Sty;使用anti-Sty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Sty原核表达,而其还获得了具有生物活性的SRY蛋白,时于深入研究Sry及其他相关基因的相互作用奠定了试验基础.
동물적성별재축목업시일항흔중요적경제성상,이포유동물적성별결정수고환결정기인Sty적계동,획득SRY단백병탐색타여기타상관인자지간적상호관계대우심입료해해인자적작용궤리화심구고효경제적성별공제방법우위중요.이소서위시험재료,통과PCR、질립중조、매절화측서등방법구건소서Sty적원핵표체재체,사용IPTG유도단백표체、SDS-PAGE전영화western blotting등방법검측중조표체재체적원핵표체.PCR、매절화측서결과일치현시성공지구건료원핵표체재체pET-21b-Sty;사용anti-Sty화anti-His특이성항체진행적western blotting시험결과험증료단백표체적정학성.해시험불부실현료포유동물성별결정인자적Sty원핵표체,이기환획득료구유생물활성적SRY단백,시우심입연구Sry급기타상관기인적상호작용전정료시험기출.
Sex of breeded animal is a very important character for stockbreeding, while for mammal, the sex determination is started by gene Sty. 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 Sty-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.