TAILIEUCHUNG - Báo cáo sinh học: " Establishment of stable Huh-7 cell lines expressing various hepatitis C virus genotype 3a protein: an in-vitro testing system for novel anti-HCV drugs"

Tuyển tập các báo cáo nghiên cứu về sinh học được đăng trên tạp chí sinh học Journal of Biology đề tài: Establishment of stable Huh-7 cell lines expressing various hepatitis C virus genotype 3a protein: an in-vitro testing system for novel anti-HCV drugs | Butt et al. Genetic Vaccines and Therapy 2011 9 12 http content 9 1 12 z GENETIC VACCINES AND THERAPY RESEARCH Open Access Establishment of stable Huh-7 cell lines expressing various hepatitis C virus genotype 3a protein an in-vitro testing system for novel anti-HCV drugs Sadia ButZ Muhammad IdreesZ Irshad-ur RehmaZ Liaqat Ali Abrar Hussain Muhammad Ali Naveed Ahmed Sana Saleem and Madiha Fayyaz Abstract Background Hepatitis C virus HCV infection is the leading cause of chronic hepatitis which progresses to hepatocellular carcinoma HCC afflicting 170 million people worldwide. HCV 3a is the most common genotype about 70 of all genotypes circulating in Pakistan. Expression of HCV individual gene of 3a would facilitate therapeutic and vaccines strategies against chronic HCV and liver Cirrhosis. The aim of the present study was the establishment of stable Huh-7 cell lines expressing structural and non structural proteins of HCV Genotype 3a Pakistani isolate obtained from chronic HCV patients. Methods Blood samples were obtained from chronic HCV-3a positive patients. HCV individual genes were amplified using PCR with gene specific primers having restriction sites. These gene amplicons were cloned in mammalian expression vector . Huh-7 cell lines were transfected with these constructed plasmids having structural or non-structural HCV genes in confluent cells with lipofectamine. Positive clones were selected with G418 and then confirmed by genome PCR. Subsequently transcription and expression of the integrated genes were demonstrated by RT-PCR sequencing and Western blot analysis. Results We successfully cloned and express five HCV-3a genes in mammalian expression vector. Results of western blot and sequencing PCR confirmed the stable expression of these five genes. Conclusion The stable cell-lines expressing HCV-3a individual genes would be a useful tool to investigate the role of various HCV proteins on HCV disease outcome and .

• Báo cáo khoa học
• Báo cáo sinh học hay
• cách trình bày báo cáo
• báo cáo sinh học
• công trình nghiên cứu sinh học
• tài liệu về sinh học