激光生物学报, 2014, 23 (4): 351, 网络出版: 2015-07-07
可诱导表达Mesp1的慢病毒载体构建及P19细胞稳定表达系的建立
Construction of Inducible Lentiviral Vector Containing Mesp1 and Establishment of a Stably Expressing Mesp1 P19 Cell Line
摘要
Mesp1属于bHLH转录因子家族, 对早期心脏中胚层的形成十分重要。为研究Mesp1在心脏发育过程中的功能, 本文构建了可诱导表达Mesp1的慢病毒载体, 转染进P19细胞, 经嘌呤霉素筛选并扩大培养后, 成功建立起经强力霉素诱导后可稳定过表达Mesp1基因的细胞系P19-Mesp1, 为研究Mesp1心脏发育相关功能提供有用的细胞研究模型。
Abstract
The Mesp1 gene encodes the basic HLH protein Mesp1 which is expressed in the mesodermal cell lineage during early gastrulation. In order to study the function of Mesp1 in heart development, inducible lentiviral vector containing Mesp1 was constructed, then it was tranfected into P19 cells to construct a stably Mesp1-expressing P19 cell lines. After screening culture by puromycin, the over expression of Mesp1 was verified by RT-PCR and Western blotting. The P19 cells line that over-express Mesp1 stably was established successfully, which provide a cell model for studying the molecular function of Mesp1 in heart development.
宋文, 高建芳, 陈辉, 邓云, 李永青, 袁婺洲, 吴秀山, 戴国, 王跃群. 可诱导表达Mesp1的慢病毒载体构建及P19细胞稳定表达系的建立[J]. 激光生物学报, 2014, 23(4): 351. SONG Wen, GAO Jianfang, CHEN Hui, DENG Yun, LI Yongqing, YUAN Wuzhou, WU Xiushan, DAI Guo*, WANG Yuequn. Construction of Inducible Lentiviral Vector Containing Mesp1 and Establishment of a Stably Expressing Mesp1 P19 Cell Line[J]. Acta Laser Biology Sinica, 2014, 23(4): 351.
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