猪Follistatin cDNA克隆及在大肠杆菌中的表达  

Porcine Follistatin cDNA Cloning and Expression in Escherichia coli

作　　者：何新[1,2] 齐冰[1,2] 何立千[3] 陈永福[4] 刘桂生[1] 陈清轩[1] 

机构地区：[1]中国科学院遗传与发育生物学研究所 [2]中国科学院研究生院,北京100049 [3]北京联合大学师范学院应用生物技术系 [4]中国农业大学农业生物技术国家重点实验室

出　　处：《生物工程学报》2006年第4期677-681,共5页Chinese Journal of Biotechnology

基　　金：北京联合大学师范学院和吉林精气神有限公司赞助。~~

摘　　要：提取猪卵巢总RNA,用RT-PCR方法克隆了猪FollistatincDNA的完整开放阅读框,长1038bp。将FollistatincDNA连接到原核表达载体pGEX-4T-3中,转化大肠杆菌BL21(DE3),以IPTG诱导,进行了GST-FS融合蛋白表达。用SDS-PAGE和Western杂交检测,结果显示在63kD处有特异性表达蛋白。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.

关 键 词：RT-PCR 猪Follistatin CDNA 原核表达 

分 类 号：Q78[生物学—分子生物学]