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| 猪Follistatin cDNA克隆及在大肠杆菌中的表达 | |
| 引用本文： | 何新 齐冰 何立千 陈永福 刘桂生 陈清轩. 猪Follistatin cDNA克隆及在大肠杆菌中的表达[J]. 生物工程学报, 2006, 22(4): 677-681 |
| 作者姓名： | 何新 齐冰 何立千 陈永福 刘桂生 陈清轩 |
| 作者单位： | 1. 中国科学院遗传与发育生物学研究所,北京,100080;中国科学院研究生院,北京,100049 2. 北京联合大学师范学院应用生物技术系,北京,100083 3. 中国农业大学农业生物技术国家重点实验室,北京,100094 4. 中国科学院遗传与发育生物学研究所,北京,100080 |
| 基金项目： | 北京联合大学校科研和教改项目 |
| 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。 |
| 关 键 词： | RT-PCR 猪Follistatin cDNA 原核表达 |
| 文章编号： | 1000-3061(2006)04-0677-05 |
| 修稿时间： | 2005-12-26 |
Porcine Follistatin cDNA Cloning and Expression in Escherichia coli |
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| HE Xin,QI Bing,HE Li-Qian,CHEN Yong-Fu,LIU Gui-Sheng,CHEN Qing-Xuan. Porcine Follistatin cDNA Cloning and Expression in Escherichia coli[J]. Chinese journal of biotechnology, 2006, 22(4): 677-681 | |
| Authors: | HE Xin QI Bing HE Li-Qian CHEN Yong-Fu LIU Gui-Sheng CHEN Qing-Xuan |
| Affiliation: | Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100080, China. |
| Abstract: | 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. |
| Keywords: | RT-PCR porcine follistatin cDNA prokaryotes expression |
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