激光生物学报, 2011, 20 (5): 681, 网络出版: 2015-10-08
稳定过表达人KLHL31基因的H9c2细胞系的建立及鉴定
Establishment and Identification of a Human KLHL31 stable-Dver Expressed H9c2 Cell Line
摘要
人类KLHL31基因是本实验室已克隆的基因, 其蛋白质含有保守的BTB和串连重复的Kelch结构域, 实验表明人类KLHL31在成体骨骼肌和心肌组织中特异表达, KLHL31蛋白的过表达可以抑制了AP-1与SRE的活性。本文拟利用阳离子聚合物转染技术将重组表达质粒pCMV-tag2B-KLHL31转染H9c2细胞, 通过G418筛选, RT-PCR和Western blotting验证, 建立稳定过表达KLHL31基因的细胞系H9c2- KLHL31, 为研究KLHL31在心脏发育及其相关功能提供有用的细胞研究模型。
Abstract
Human KLHL31 gene has been cloned previously in our lab, and the protein product contains a BTB domain and a Keleh-repeat domain. It was reported that KLHL31 especially expressed in skeletal and cardiac muscles of adult tissues, In MAPK signal transduction pathways, overexpression of KLHL31 inhibited the transcriptional activities of AP-1 and SRE. To construct a stable KLHL31-expressed H9c2 cell lines. The fusion expression plasmid pCMV-tag2B-KLHL31 was transfected into H9c2 cell by cationic polymer. After screening culture by G418, the over expression of KLHL31 was verified by RT-PCR and Western blotting. The H9c2 cells line that over-expressing KLHL31 stably was established successfully, which provide a cell model for our study the molecular function of KLHL31 in heart development.
周军媚, 叶湘漓, 廖四芳, 刘明, 吴秀山, 王跃群. 稳定过表达人KLHL31基因的H9c2细胞系的建立及鉴定[J]. 激光生物学报, 2011, 20(5): 681. ZHOU Junmei, YE Xiangli, LIAO Sifang, LIU Ming, WU Xiushan, WANG Yuequn. Establishment and Identification of a Human KLHL31 stable-Dver Expressed H9c2 Cell Line[J]. Acta Laser Biology Sinica, 2011, 20(5): 681.
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