Construction and expression of non-structural protein gene 3-4b of HCV 1b based on the adenoassociated virus vector
10.3760/cma.j.issn.1003-9279.2013.02.016
- VernacularTitle:1b型丙型肝炎病毒NS3-4b重组腺相关病毒载体的构建及表达
- Keywords:
Hepatitis C;
Hepatitis viruses;
Adenoviridae
- From:
Chinese Journal of Experimental and Clinical Virology
2013;27(2):132-134
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone 1b type of HCV NS3-4b Gene and express in HEK 293 cells,lay the foundation for further study of the HCV NS3-4b recombinant adeno-associated virus vaccine and its dendritic cell vaccine.Methods HCV 1b patients' serum was collected,and full length NS3-4b segment was amplified by RT-PCR and cloned into adeno-associated virus' expression vector pAAV.CMV.EGFP in order to express in HEK 293 cells.At last,it was validated whether express or not by Western Blot.Results The 1b type gene NS3-4b were amplified and consistent to the expected size(2838 bp),the recombinant plasmid has been confirmed its successful restructured by double enzyme and sequencing,at last,Western Blot map can see objective protein expression after it transfect HEK 293 cells.Conclusion The adenoassocisted virus recombination HCV NS3-4b plasmid have successfully constructed and it can express in eukaryotic cells.
