[Establishment of COL1A1-EGFP stably transfected ROS17/2.8 cell lines].
OBJECTIVE
To obtain ROS17/2.8 cell lines which were stably expressing EGFP reporter gene drived by COL1A1 promoter.


METHODS
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.


RESULTS
The COL1A1-EGFP stably transfected cell lines were established.


CONCLUSIONS
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.
Dai
Zhong-quan
ZQ
Institute of Space Medico-Engineering, Beijing, China.
Li
Ying-hui
YH
Ding
Bai
B
Zhang
Yu-guo
YG
Liu
Peng-peng
PP
D000469:Alkaline Phosphatase / Q000235:genetics; D000818:Animals; D002460:Cell Line; D024042:Collagen Type I / Q000235:genetics*; D000090602:Collagen Type I, alpha 1 Chain; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics*; D007150:Immunohistochemistry; D010006:Osteoblasts; D012516:Osteosarcoma / Q000235:genetics; D016133:Polymerase Chain Reaction; D051381:Rats; D018474:Weightlessness Simulation
D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D000469:Alkaline Phosphatase