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| TES基因C端的分段克隆与表达 | |
| 引用本文： | 冯波,曾虹燕,钟英丽,张健. TES基因C端的分段克隆与表达[J]. 广西农业生物科学, 2007, 26(3): 205-209 |
| 作者姓名： | 冯波 曾虹燕 钟英丽 张健 |
| 作者单位： | 1. 湖南师范大学,生命科学院,湖南,长沙,410081;湘潭大学,化工学院,湖南,湘潭,411105 2. 湘潭大学,化工学院,湖南,湘潭,411105 3. 湖南师范大学,生命科学院,湖南,长沙,410081 |
| 基金项目： | 国家自然科学基金项目(30470945)，湖南省教育厅项目(06C816) |
| 摘 要： | 为了在大肠杆菌中分段表达人类TES基因C端3个顺次排列的LIM结构域,利用PCR技术从含有TES基因全长编码序列的质粒pCMV-HA-TES中扩增出这3个LIM domain基因片段,分别重组入pGEM-T载体,再经酶切后,将获得的目的片段亚克隆入原核表达载体pQE-N1进行融合表达。酶切鉴定及测序结果表明,扩增产物与已知基因序列一致,成功构建了3个目的片段的pQE-N1表达质粒;SDS-PAGE结果显示,IPTG诱导表达出与预期分子质量大小相同的3个融合蛋白。 |
| 关 键 词： | TES基因 LIMdomain 亚克隆 蛋白表达 |
| 文章编号： | 1008-3464（2007）03-0205-05 |
| 收稿时间： | 2006-11-14 |
| 修稿时间： | 2007-01-04 |
Cloning and expression of truncated TES C-terminus |
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| FENG Bo,ZENG Hong-yan,ZHONG Ying-li,ZHANG Jian. Cloning and expression of truncated TES C-terminus[J]. Journal of Guangxi Agricultural and Biological Science, 2007, 26(3): 205-209 | |
| Authors: | FENG Bo ZENG Hong-yan ZHONG Ying-li ZHANG Jian |
| Affiliation: | 1.College of Life Science, Hunan Normal University, Changsha 410081, China; 2.College of Chemical Engineering, Xiangtan University, Xiangtan 411105, China |
| Abstract: | In order to express three tandem LIM domains in the C-terminus of TES individuallyin Escherichia coli BL21,three gene fragments encoding corresponding LIM domains in TESC-terminus were amplified from pCMV-HA-TES plasmid containing the full length of TES coding sequence by PCR and cloned into pGEM-T vector respectively.The target gene fragments obtained by digesting the recombinant plasmids with restriction endonuclease were subcloned into prokaryotic expression vector pQE-N1 for fusion expression.The results of restriction endonuclease digestion and sequence analysis indicated that three recombinant plasmids for expression of corresponding LIM domains in TES C-terminus were successfully constructed.The fusion proteins with the molecular weights as prospected were separated by SDS-PAGE gel after induced expression. |
| Keywords: | TES gene LIM domain subcloning protein expression |
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