Control of Thrombopoietin Expression by Doxycycline in CHO Cells
- Author:
Shuang FU
1
,
2
;
Xudong WEI
;
Kang CHENG
;
Aili LU
;
Shengwu WANG
;
Debing WANG
Author Information
1. Institute of Hematology, People's Hospital, Beijing Medical University, Beijing 100044, China
2. Department of Bichemistry and Molecular Biology, Beijing Medicine University, Beijing 100083, China.
- Publication Type:Journal Article
- From:
Journal of Experimental Hematology
2000;8(1):10-13
- CountryChina
- Language:Chinese
-
Abstract:
To control thrombopoietin (TPO) expression by doxycycline in CHO cells, Tet-On gene expression system was used. Recombinant plasmid pTRE-TPO was successfully constructed. pTRE-TPO and pTK-Hyg were cotransfected into CHO-Tet-On cells. Cells resistant to hygromycin were cloned, high expression and low background clone was selected, and named as CHO-Tet-On-TPO. There was no significant TPO expression in the absence of doxycycline, the TPO level in the cell culture supernatant was 0.1 micro g/L. After exposure to 2 mg/L doxycycline, TPO expression was greatly increased, the TPO level in the cell culture supernatant reached 10.8 micro g/L. Tet-On gene expression system is a ready access to the tight-regulated and high-level gene expression system. It is likely to provide a safe and regulatable way for TPO gene therapy.
