Dai Zhong-quan, Li Ying-hui, Ding Bai, Zhang Yu-guo, Liu Peng-peng
Institute of Space Medico-Engineering, Beijing, China.
Space Med Med Eng (Beijing). 2004 Apr;17(2):107-10.
To obtain ROS17/2.8 cell lines which were stably expressing EGFP reporter gene drived by COL1A1 promoter.
A 3.6 Kb COL1A1 promoter from rat was cloned into pMD-18-T vector by PCR. This amplified promoter vector was digested to get several different length fragments which were then fused with EGFP reporter gene to construct eukaryotic expression vectors. ROS17/2.8 cell was stably transfected with these vectors by LipofectAMINE(TM) and selected by G418.
The COL1A1-EGFP stably transfected cell lines were established.
The cell lines will be useful for studying the effects of microgravity on the activity of COL1A1 promoter and expression of gene related with bone form.
获得由COL1A1启动子驱动稳定表达EGFP报告基因的ROS17/2.8细胞系。
通过PCR将大鼠的3.6 Kb COL1A1启动子克隆到pMD-18-T载体中。将扩增的启动子载体进行酶切以获得几个不同长度的片段，然后将这些片段与EGFP报告基因融合以构建真核表达载体。通过脂质体转染法用这些载体对ROS17/2.8细胞进行稳定转染，并用G418进行筛选。
建立了COL1A1-EGFP稳定转染的细胞系。
该细胞系将有助于研究微重力对COL1A1启动子活性及骨形成相关基因表达的影响。
