Objective: To investigate four expression vectors carrying enhanced green fluorescent protein (EGFP) genes, which under the control of different HBV promoters, were detected and compared as to their expressions in hep...Objective: To investigate four expression vectors carrying enhanced green fluorescent protein (EGFP) genes, which under the control of different HBV promoters, were detected and compared as to their expressions in hepatocytes and non- hepatic cell lines. Methods: Four HBV promoters (including enhancers) were amplified by PCR, subcloned into the expression vector pEGFP-1. The four recombinants controlled by HBV promoters were analyzed and identified by restriction enzymes and sequencing. After transfection into human hepatoma cell lines and non-liver cells, transfection efficiency was measured by EGFP expression through fluorescence microscopy and analyzed with fluorescence activated cell sorter (FACS). Results: All target fragments were separately obtained and successfully cloned into the expression vector. The transfection results showed that in hepatoma cells, the expression of EGFP was more obvious than it was in non-hepatic cells. Among the four promoters, S gene promoter presented the strongest transfection efficiency. Conclusion: Our findings indicate that HBV promoters could lead specific expression in hepatocytes, different promoters had different outcomes, which might be a potential for the gene therapy of liver diseases.展开更多
The anti-hepatitis B virus(HBV)effects and its mechanisms of the ethanol extracts of Hypericum perforatum L.(EHP)in vitro were explored.HepG2 2.2.15 cells,a stable HBV-producing cell line,were cultured as the model sy...The anti-hepatitis B virus(HBV)effects and its mechanisms of the ethanol extracts of Hypericum perforatum L.(EHP)in vitro were explored.HepG2 2.2.15 cells,a stable HBV-producing cell line,were cultured as the model system to observe the anti-HBV effect.The viral antigens of cellular secretion,HBsAg and HBeAg,were determined by enzyme linked immunosorbent assay(ELISA).The quantity of HBV-DNA released in the supernatant was assayed by real-time PCR.In order to understand the mechanisms of the suppression of H...展开更多
基金Supported by a grant form the National Nature Science Foundation of China (30330680).
文摘Objective: To investigate four expression vectors carrying enhanced green fluorescent protein (EGFP) genes, which under the control of different HBV promoters, were detected and compared as to their expressions in hepatocytes and non- hepatic cell lines. Methods: Four HBV promoters (including enhancers) were amplified by PCR, subcloned into the expression vector pEGFP-1. The four recombinants controlled by HBV promoters were analyzed and identified by restriction enzymes and sequencing. After transfection into human hepatoma cell lines and non-liver cells, transfection efficiency was measured by EGFP expression through fluorescence microscopy and analyzed with fluorescence activated cell sorter (FACS). Results: All target fragments were separately obtained and successfully cloned into the expression vector. The transfection results showed that in hepatoma cells, the expression of EGFP was more obvious than it was in non-hepatic cells. Among the four promoters, S gene promoter presented the strongest transfection efficiency. Conclusion: Our findings indicate that HBV promoters could lead specific expression in hepatocytes, different promoters had different outcomes, which might be a potential for the gene therapy of liver diseases.
文摘The anti-hepatitis B virus(HBV)effects and its mechanisms of the ethanol extracts of Hypericum perforatum L.(EHP)in vitro were explored.HepG2 2.2.15 cells,a stable HBV-producing cell line,were cultured as the model system to observe the anti-HBV effect.The viral antigens of cellular secretion,HBsAg and HBeAg,were determined by enzyme linked immunosorbent assay(ELISA).The quantity of HBV-DNA released in the supernatant was assayed by real-time PCR.In order to understand the mechanisms of the suppression of H...
