CTGF靶向RNA干扰重组体的构建及序列分析
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主余华, 张春清, 赵幼安, 褚衍六, 孙成刚
结缔组织生长因子;RNA干扰;短发夹RNA;质粒,主余华,张春清,孙成刚,赵幼安,褚衍六,通讯作者:,电话:,收稿日期:,接受日期:,Construc
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主余华, 张春清, 孙成刚, 山东大学山东省立医院消化内科 山东省济南市 250021
赵幼安, 褚衍六, 山东大学齐鲁医院消化内科 山东省济南市 250012
山东省卫生厅青年基金资助项目, No. 2005-25
通讯作者: 张春清, 250021, 山东省济南市, 山东省立医院消化内科. zyh6698@yahoo.com.cn
电话: 0531-85186350
收稿日期: 2005-10-13 接受日期: 2005-11-15
Construction and sequence analysis of connective tissue growth factor targeted recombinant plasmids by RNA interference
Yu-Hua Zhu, Chun-Qing Zhang, You-An Zhao, Yan-Liu Chu, Cheng-Gang Sun
Yu-Hua Zhu, Chun-Qing Zhang, Cheng-Gang Sun, Department of Gastroenterology, Shandong Provincial Hospital of Shandong University, Ji’nan 250021, Shandong Province, China
You-An Zhao, Yan-Liu Chu, Department of Gastroenterology, Shandong Qilu Hospital of Shandong University, Ji’nan 250021, Shandong Province, China
Supported by the Fund for Young Scholars from Shandong Provincial Health Department, No. 2005-25
Correspondence to: Chun-Qing Zhang, Department of Gastro-enterology, Shandong Provincial Hospital of Shandong University, Ji’nan 250021, Shandong Province, China. zyh6698@yahoo.com.cn
Received: 2005-10-13 Accepted: 2005-11-15
Abstract
AIM: To construct the recombinant plasmids expressing connective tissue growth factor (CTGF) short hairpin RNA (shRNA) by pEGFP plasmid vector.
METHODS: Three pairs of two DNA sequences containing small hairpin structure were designed and synthesized respectively, and then were formed into complementary chains by annealing. The obtained products were inserted into plasmid vector pEGFP containing U6 promoter. Then the recombinant plasmids were transformed into DH5α strain. Finally the plasmids that identified by restriction enzyme were used for sequence analysis.
RESULTS: The CTGF shRNA expression frames were successfully inserted into the plasmid vector pEGFP, and the shRNA coding sequences of the 3 obtained recombinant plasmids were consistent with the designed fragments ......
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