[The influences of Tet system on activity and tissue specificity of HBV Cp.].
OBJECTIVE
Analyze the effect of Tetracycline-controlled system on Hepatitis B virus core promoter (Cp) activity and tissue specificity.


METHODS
The 7 Teto sequence was amplified from plasmid pTL-8 using polymerase chain reaction (PCR) and cloned into T vector pMD19-T Simple. After sequencing, the 7 teto sequence was subcloned into upstream of Cp in pGL3-Basic/Cp. In order to observe the effect of Tetracycline-controlled system on Cp activity and tissue specificity, the plasmid pTL-8 which luciferase encoding sequence was excised and pGL3-Basic/7t/Cp were cotransfected into different tissue-derived cell lines including HepG2, Hela, COS-7, MDA-MB-231 and HT-29. The dual luciferase activities were determined by Dual Luciferase Report (DLR) assay system.


RESULTS
Have successfully constructed plasmid pGL3-Basic/7t/Cp. The transcriptional activity of Cp was increased significantly after the teto sequence was cloned upstream of Cp.


CONCLUSIONS
The transcriptional activity of Cp is augmented significantly by Tetracycline-controlled system. But the tissue specificity of Cp lost at the same time.
Lei
Jun-Chuan
JC
Department of Etiology, School of Basic Medicine, Xi'an 710032, China.
Gong
Wei-Dong
WD
Zhao
Ya
Y
Jiang
He
H
Liu
Zhong-Xiang
ZX
Yi
Jun
J
D005822:Genetic Vectors; D006515:Hepatitis B virus / Q000378:metabolism; D006801:Humans; D008156:Luciferases / Q000235:genetics; D009928:Organ Specificity; D010957:Plasmids; D011401:Promoter Regions, Genetic
D008156:Luciferases