| In order to approach the effect that RNAi restrains proliferation of Bombyx mori nucleopolyhedrovirus virus(BmNPV), BmN cells transfected with transgenic vector pigA3-LEF-Neo containing lef-1 dsRNA transient expression box and stable transformation cell line screened with G418(750-800μg/mL) were procured. Virus infection test showed that the infection ratio of the stable transformation cells was lower than that of normal cells by 53% ;the quantity of polyhedron from transformation cells were 2/3 time of that of normal cells and the number of liberation virus from cell culture's supernatant decreased 10-20 times. The result indicated that the virus' proliferation in the transformation cells was depressed significantly. That the transcriptional level of virus' lef-1 in the transformation cells was only 2/5-3/5 of that in the normal cells suggested that the transcribed lef-1 dsRNA in transformation cells inhibited the expression of virus' lef-1. Inverse PCR was used to locate the site of exogenous DNA fragment insertion in genome, of which the result showed that in the transformation cells, exogenous DNA could randomly or preferringly based on piggyBac's special transposition site TTAA integrate into the cellular genome.By sperm-mediated gene transferring and particle gun bombarding, the transgenic vector pigA3-LEF-Neo was carried into the eggs of silkworm.the larvae out of eggs were bred in natural environment without disease prevention measure, in the end, five survivals pupa all showed different extent fluorescence intensity screened with neo and gfp. By PCR identifying,the result confirmed that what the fluorescence silkworms procured were transgenic silkworms.by initial screening,the result showed that the fluorescence silkworms of G0 depressed BmNPV significantly.four kinds of transgenic silkworms of G1 were procured by array hybridization of G0's fluorescence silkworms,that established foundation for gaining transgenic silkworms which depressed BmNPV in the end.
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