He Xin, Qi Bing, He Li-Qian, Chen Yong-Fu, Liu Gui-Sheng, Chen Qing-Xuan
Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100080, China.
Sheng Wu Gong Cheng Xue Bao. 2006 Jul;22(4):677-81.
The total RNA was extracted from porcine ovary. Porcine Follistatin cDNA was cloned by RT-PCR. Complete porcine follistatin cDNA coding sequences are presented including 1038 bp of open reading frame. The purified porcine follistatin cDNA was inserted into pGEX-4T-3 vector to construct the prokaryotic fusion protein expression vector. The recombinant expression plasmid was transformed into BL21 (DE3) and expression was induced by IPTG. Protein products were detected by SDS-PAGE and confirmed by Western blotting analysis, which showed that the yield of the Follistatin cDNA was a 63kD protein expression vector. Follistatin protein was expressed in the form of glutathione-S-transferase (GST) fusion protein in E. coli.
从猪卵巢中提取总RNA。通过逆转录聚合酶链反应（RT-PCR）克隆猪卵泡抑素cDNA。呈现了完整的猪卵泡抑素cDNA编码序列，包括1038 bp的开放阅读框。将纯化的猪卵泡抑素cDNA插入pGEX-4T-3载体中，构建原核融合蛋白表达载体。将重组表达质粒转化到BL21(DE3)中，并用异丙基-β-D-硫代半乳糖苷（IPTG）诱导表达。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）检测蛋白产物，并通过蛋白质免疫印迹分析进行确认，结果表明卵泡抑素cDNA的产量为一种63kD的蛋白表达载体。卵泡抑素蛋白在大肠杆菌中以谷胱甘肽-S-转移酶（GST）融合蛋白的形式表达。
