Tian Chen, Hai-Xia Sun, Wei-Bin Qin, Feng-Qin Zhu, Xue-Ling Li, Hong Cao, Qi-Huan Xu, Gang Li
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[Construction and expression of non-structural protein gene 3-4b of HCV 1b based on the adenoassociated virus vector].
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 adeno-associsted virus recombination HCV NS3-4b plasmid have successfully constructed and it can express in eukaryotic cells.
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