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ISSN
1672-9145
Acta Biochim Biophys Sin
2004, 36(10): 687–694
CN 31-1940/Q
Temporal and Tight Hepatitis C Virus Gene Activation in Cultured
Human Hepatoma Cells Mediated by a Cell-Permeable Cre Recombinase
Dong XIAO*, Kang XU1,
Ying YUE, Zhong-Min GUO, Bing HUANG, Xin-Yan DENG, Huan TANG2, and Xi-Gu CHEN*
Center of
Experimental Animals, Zhongshan University, Guangzhou 510080, China;
1The first affiliated
hospital of Sun Yat-sen University, Zhongshan University, Guangzhou 510080,
China;
2The Third Military
Medical University, Chongqing 400010, China
Abstract Conditional gene
expression has greatly facilitated the examination of the functions of
particular gene products. Using the Cre/lox P switching
expression system, we plan to develop efficient conditional transgene activation of hepatitis C virus core protein (HCV-C) cDNA
(nucleotide 342914) in the transgenic mice to overcome �immune tolerance formed during the
embryonic period and �immune escape against hepatitis virus antigen in our project. To use
this system in vivo, the dormant transgenic construct, i.e.,
pApoE-SCS-EGFP-HCV-C, was generated using techniques of standard molecular biology.
The liver-specific human apoE promoter was here used to target
expression of genes of interest (EGFP and HCV-C) to murine liver. Prior to
generating the transgenic mice, the availability of Cre/lox P system and
construct functionality were successfully verified by a cell-free recombination
system and via checking the expression of EGFP and HCV-C in the human hepatoma
cells at the mRNA and protein levels. These results suggest that the Cre/lox
P system could tightly control expression of EGFP and HCV-C in vitro,
which laid a solid foundation to conditionally activate expression of target
gene(s) in transgenic mice by Cre-mediated site-specific recombination.
Key words hepatitis C virus; Cre/lox
P switching expression system; cell-free and intracellular recombination
systems; cell-permeable Cre recombinase; EGFP; human hepatoma cells
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Received: March 25, 2004 Accepted: September 10,
2004
This work was supported by the grants from the National Natural
Science Foundation of China (No. 30271177, No. 39870676), Guangdong Province
Natural Science Foundation of China (No. 021903) and Postdoctoral Fellowship
Foundation of China (Series 29)
*Corresponding author: Tel, 86-20-87331393; Fax, 86-20-87331230;
E-mail, [email protected]