Construction and function of recombinant AcMNPV with double copies of v-cath gene |
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Authors: | Deli Liu Huazhong Xiao Yipeng Qi Lunguang Yao |
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Institution: | (1) Institute of Virology, Wuhan University, 430072 Wuhan, China |
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Abstract: | Two recombinant baculoviruses, dciAcMNPV and dcdAcMNPV in which another copy of the v-cath gene controlled by ie1 promoter and polh promoter was inserted, were respectively constructed by the Bac-to-Bac system. The expression of the v-cath gene of the recombinant baculoviruses in Sf9 cells at different phases was investigated by SDSPAGE and Western blot. The
results showed that only recombinant virus dciAcMNPV containing late gene v-cath driven by early gene promoter could express V-CATH protein, cathepsin encoded by virus genome, 12 h post-infection and dcdAcMNPV
containing late gene v-cath driven by late and very late gene promoters could express more V-CATH protein. Negative control ncAcMNPV, a mutant deleted
vcath gene, could not express V-CATH protein at all. The Spodopera exigua larvae were infected with viruses respectively and the results showed that the toxicity was as follows: dcdAcMNPV>dciAcMNPV>wtAcMNPV>ncAcMNPV.
The toxicity of recombinant viruses and the characters of dead larvae showed that the v -cath gene was relative to viral toxicity and host liquefaction. Recombinant baculovirus dcdAcMNPV might be used as a new kind
of safe viral-pesticide, because of its high toxicity obtained by adding another gene copy and changing the expression level
of its own gene relative to virulence. |
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Keywords: | v -cath gene Western blot Bac-to-Bac system recombinant baculovirus |
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