VEGF-C基因靶向RNA干扰重组表达载体的构建和表达
胃癌;血管内皮生长因子-C;重组质粒;RNA干扰,周慧聪,李继昌,亢春彦,张艳,通讯作者:,ConstructionandexpressionofsmallinterferingRNAvectortargetingonvascula
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周慧聪, 李继昌, 郑州大学第一附属医院消化内科 河南省郑州市 450052
亢春彦, 河南省职工医学院病理教研室 河南省郑州市 450052
张艳, 郑州大学基础医学院药理教研室 河南省郑州市 450052
通讯作者: 周慧聪, 450052, 河南省郑州市, 郑州大学第一附属医院消化内科. hczhouar@163.com
电话: 0371-66862182
收稿日期: 2006-11-28 接受日期: 2007-01-27
Construction and expression of small interfering RNA vector targeting on vascular endothelial growth factor-C
Hui-Cong Zhou, Chun-Yan Kang, Yan Zhang, Ji-Chang Li
Hui-Cong Zhou, Ji-Chang Li,Department of Gastroenterology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, He’nan Province, China
Chun-Yan Kang,Department of Pathology, He’nan Medical College for Staff and Workers, Zhengzhou 450052, He’nan Province, China
Yan Zhang,Department of Pharmacology, Basic Medical Sciences of Zhengzhou University, Zhengzhou 450052, He’nan Province, China
Correspondence to:Hui-Cong Zhou, Department of Gastroenterology, the First Affiliated Hospital of Zhengzhou University, Zhenzhou 450052, He’nan Province, China. hczhouar@163.com
Received:2006-11-28 Accepted:2007-01-27
Abstract
AIM: To clone the recombinant plasmid affecting vascular endothelial growth factor-C (VEGF-C) gene transcription by RNA interference and observe VEGF-C expression in human gastric cancer cell.
METHODS: Two DNA sequences containing small hairpin structure were designed and synthesized. The complement form was obtained by annealing and then cloned into vector pSilencer3.1-H1. The recombinant plasmid was transformed into strain DH5α. After amplification and purification, the target plasmid, identified by restriction enzyme, was obtained sequenced. Finally, gastric carcinoma cell line SGC-7901 was transfected with the plasmid, and the protein expression of VEGF-C was detected by Western blot.
RESULTS: After enzyme digestion and sequencing, the VEGF-C siRNA expression vector was successfully constructed ......
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