干扰Fas受体表达的串联shRNA表达载体的构建
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刘明社, 王 兰, 赵中夫, 张国英, 张 芸,
Fas;RNAi;shRNA;串联质粒刘明社,王兰,赵中夫,张国英,张芸,封江南.干扰Fas受体表达的串联shRNA表达载体的构建. 世界华人消化杂志2006;14(22)2174-217
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刘明社, 赵中夫, 张国英, 张芸, 长治医学院肝病研究所, 山西省长治市 046000
王兰, 山西大学生命科学与技术学院 山西省太原市 030006
封江南, 武汉市晶赛生物工程技术有限公司 湖北省武汉市430074山西省自然基金资助项目, No. 20051114
通讯作者: 赵中夫, 046000, 山西省长治市解放东街161号, 长治医学院肝病研究所. zhaozf_1226@163.com
电话: 0355-3012335
收稿日期: 2006-03-26 接受日期: 2006-06-14
Construction of recombinant plasmid series with two Fas-targeted short hairpin RNAs
Ming-She Liu, Lan Wang, Zhong-Fu Zhao, Guo-Ying Zhang, Yun Zhang, Jiang-Nan Feng
Ming-She Liu, Zhong-Fu Zhao, Guo-Ying Zhang, Yun Zhang, Institute of Hepatology, Changzhi Medical College, Changzhi 046000, Shanxi Province, China
Lan Wang, College of Life Science and Technology, Shanxi University, Taiyuan 030006, Shanxi Province, China
Jiang-Nan Feng, Wuhan Genesil Biotechnology Corporation Limited, Wuhan 430074, Hubei Province, China
Supported by the Natural Science Foundation of Shanxi Province of China, No. 20051114
Correspondence to: Zhong-Fu Zhao, Institute of Hepatology, Changzhi Medical College, 161 Jiefang East Street, Changzhi 046000, Shanxi Province, China. zhaozf_1226@163.com
Received: 2006-03-26 Accepted: 2006-06-14
Abstract
AIM: To construct the recombinant plasmid expressing two Fas-targeted short hairpin RNA (shRNA) by pEGFP6-1 and pGenesil-1 plasmids vector.
METHODS: Two pairs of DNA sequences were designed, and then synthesized into complementary chains by annealing, respectively. Then the obtained products containing short hairpin structure were inserted into plasmid vector pEGFP6-1/pGenesil-1 with U6 promoter. The recombinant plasmids were transformed into Escherichia coli strain DH5a for screening and amplifying. The sequence analysis of the plasmids identified by restriction enzyme were carried out. The two plasmids were first digested with SacⅠ, and then with SalⅠ. A long segment from pEGFP6-1-siFas1 and a short segment from pGenesil-1-siFas2 were reclaimed, and connected with T4 DNA Ligase ......
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