摘要
目的用大肠杆菌表达重组人促甲状腺素受体膜外区(hTSHR-ECD)蛋白,为hTSHR膜外区的研究提供工具。方法从人甲状腺组织中提取总RNA,RT-PCR分离得到TSHR膜外区cDNA,双酶切后插入原核表达载体pRSETC中,进行序列分析;再转化到表达菌BL21中表达、纯化、Western blotting分析。结果经SDS-PAGE电泳有一条明显的蛋白带,与预期分子量相符,并用Western blot证实;纯化后,获得单一条带的靶蛋白。结论成功获得高纯度人TSHR膜外区蛋白。
Objective To express and purify human thyrotropin receptor ectodomain （hTSHR-ECD） for further functional study. Methods Total RNA was isolated from human thyroid, hTSHR-ECD cDNA was obtained with RT-PCR technique, subcloned into the plasmid pRSET C ,and expressed in a soluble form in E. Coli B121. Results There was an obvious band in SDS-PAGE electrophoresis, whose molecular weight was equal to that we designed. Western blot proved the expressed product was hTSHR-ECD protein. Conclusion High pure target protein,hTSHR-ECD ,was obtained.
出处
《广东药学院学报》
CAS
2009年第5期519-521,共3页
Academic Journal of Guangdong College of Pharmacy
