[Transfer and Expression of Recombinant Human Thrombopoietin Gene in COS-7 Cells and Mice In Vivo]
In order to investigate the expression of recombinant TPO gene in COS-7 cells and in vivo of the mouse model, eukaryotic expressing plasmid pcd2/TPO with human TPO cDNA was constructed with DNA recombinant techniques. The plasmid pcd2/TPO was transiently transfected into the COS-7 cells by means of lipofection, the naked pcd2/TPO plasmid was injected into the skeletal muscle of mice with electric pulses. RT-PCR and ELISA methods were used to detect the TPO expression of the transfected COS-7 cells, both showed high level expression. The MTT test showed the expressed TPO had proliferative activity to TPO-dependent cell line. High efficiency of gene transfer in transgenic mice was also observed by RT-PCR and immunohistochemical methods. The serum TPO level [(1 185 +/- 264) ng/L] in transgenic mice was quite different compared with the normal mice [(250 +/- 76) ng/L]. All these results provided solid foundations for the research of TPO gene therapy in the future.
Zang
Wei-Ping
WP
Institute of Hematology, People's Hospital, Peking University, Beijing 100044, China; Institute of Cardiovascular Research, Peking University, Beijing 100083, China.
Wei
Xu-Dong
XD
Fu
Shuang
S
Wang
Shen-Wu
SW
Tang
Jian
J
Wang
De-Bing
DB