[Identification and evaluation promoter sequence and the transcription activation of Hcbp6 interaction with core protein of hepatitis C virus].
OBJECTIVE
To clarify the expression and regulation mechanism of the new target gene (human hepatitis C virus binding protein 6, Hcbp6) interaction with the core protein of hepatitis C virus (HCV).


METHODS
Referring to the prediction online of the promoter region, 3256 base pairs (bp) upstream and downstream of the translation start site were selected as 5 putative promoter sequences which were amplified from the hepatoblastoma cell line-HepG2 cell genomic DNA by polymerase chain reaction (PCR). The amplified products were cloned into a pCAT3 vector. The HepG2 and NIH3T3 cell lines were transfected by pCAT3-Hcbp6-promoters. The CAT activity was detected using an enzyme-linked immunoassay (ELISA) kit.


RESULTS
We found that 2 kinds of pCAT3-Hcbp6-promoters (1066 and 240) could direct the reporter gene expression. The expression of CAT was 3.1 times and 6.4 times higher than that of the pCAT3-basic control vector.


CONCLUSIONS
These results indicated that pCAT3-Hcbp6-promoters have promoter activity.
Wang
Lin
L
Center for Viral Hepatitis Research, Institute of Infectious Diseases, 302 Hospital of PLA, Beijing 100039, China. wl@genetherapy.com.cn
Cheng
Jun
J
Li
Ke
K
Hong
Yuan
Y
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D015967:Gene Expression Regulation, Viral; D017930:Genes, Reporter / Q000235:genetics; D006406:Hematoma / Q000473:pathology; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D015533:Transcriptional Activation; D014162:Transfection; D014407:Tumor Cells, Cultured; D014758:Viral Core Proteins / Q000235:genetics*
D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus