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- W1177210740 abstract "Grouper is one of the most valuable commercial marine fish and viral nervous necrosis(VNN) is the most harmful disease of this fish.Capsid protein(major capsid protein,MCP) of NNV is a major pathogenic factor and currently there is no effective drug available for its treatment.This paper is intended to provide a preliminary study of the RNAi control technology for NNV.The grouper nervous necrosis virus MCP gene was cloned and eukaryotic expression vector pEGFP-MCP was constructed with a green fluorescent protein(EGFP) fusion gene and the MCP gene.Three pairs of siRNA(NNV-001,NNV-002,NNV-003) against the gene sequence designed,were transfected into FHM cells(muscle cells of fathead minnow)with Lipofectamine2000 in order to detect EGFP expression and comparison of transfection method,dose and efficiency.The results showed that the transfection efficiency with serum-free medium after 4-6 h is higher than the rate of transfection with mixture medium.100 ng was found to be the most appropriate amount when the plasmid was transfected with 50,70,100 and 150 ng respectively.In the cotransfection of siRNA and pEGFP,all of the three pairs of siRNA sequences had interference effects on EGFP expression and pair 002 was found to be the best.Better interference effects were obtained at the siRNA concentration 200 nmol/L." @default.
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- W1177210740 date "2011-01-01" @default.
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- W1177210740 title "Preliminary study on RNA interference against red-spotted grouper nervous necrosis virus-optimization of transfection condition and comparison of interference effect based on FHM cell." @default.
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