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Construction and function of recombinant AcMNPV with double copies of v-cath gene
  • 时间:0
  • 分类:Q78[生物学—分子生物学]
  • 作者机构:Wuhan Univ, Inst Virol, Wuhan 430072, Peoples R China
  • 相关基金:This work was supported by the National Natural Science Foundation of China (Grant No. 39870041) ; Hubei Provincial Natural Science Foundation (Grant No. 971048).
中文摘要:

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 baculovi-ruses in Sf9 cells at different phases was investigated by SDS-PAGE and Western blot. The results showed that only re-combinant virus dciAcMNPV containing late gene v-cath driven by early gene promoter could express V-CATH pro-tein, 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 v-cath gene, could not express V-CATH protein at all. The Spodopera exigua larvae were infected with viruses respec-tively 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 show

英文摘要:

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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