標題: Development of Single-Vector Tet-on Inducible Systems with High Sensitivity to Doxycycline
作者: Chao, Jiun-Shuan
Chao, Chun-Chieh
Chang, Chu-Li
Chiu, Yi-Rong
Yuan, Chiun-Jye
生物科技學系
分子醫學與生物工程研究所
Department of Biological Science and Technology
Institute of Molecular Medicine and Bioengineering
關鍵字: Tetracycline;Tet-on inducible system;Single-vector;Transgenic cell;CMV;MMTV
公開日期: 1-七月-2012
摘要: Single-vector Tet-on systems were developed to enable the tight regulation of transgenes in mammalian cells with a low dosage of doxycycline. Both the regulatory and the responsive units were integrated in a single vector and separated by a short DNA segment (214 bp). In the developed single-vector Tet-on systems, a high level of expression of the transgene can be induced by doxycycline at a concentration of as low as 1 ng/ml, which is 500-1,000 times lower than that usually utilized in other Tet-on systems. The single-vector Tet-on system developed here exhibited 3.5-10.8 times greater inducibility of the transgene in response to doxycycline than did a dual-vector system from a commercial source. Further studies indicate that the basal activity of Tet-on systems depends greatly on the strength of the promoter that controls the transactivator. The basal activity of Tet-on systems was high when the transactivator that was directed by the human cytomegalovirus promoter, and it was almost undetectable when the transactivator was placed under the control of a moderate strength mouse mammary tumor virus promoter. Moreover, the introduction of selectable markers allows the developed single-vector Tet-on systems to facilitate the generation of conditional transgenic cells and animals with high inducibility, low basal activity and detrimental effects of the long-term administration of doxycycline.
URI: http://hdl.handle.net/11536/16256
ISSN: 1073-6085
期刊: MOLECULAR BIOTECHNOLOGY
Volume: 51
Issue: 3
結束頁: 240
顯示於類別:期刊論文


文件中的檔案:

  1. 000304404900005.pdf