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摘要
In this study we aimed to identify the proteinprotein interactions between Bombyx mori midgut and the bidensovirus BmDNV-Z via a yeast two-hybrid (Y2H) system. To achieve this we constructed a Gal4 activation domain fusion library that expresses the host genes and Gal4 DNA binding domain fusion bait vectors that express BmDNV-Z genes. Y2H assay revealed 15 potential interactions between host and viral proteins. To verify the interactions, we modified and reconstructed a pair of bimolecular fluorescence complementation (BiFC) vectors and achieved the co-expressions of the candidate host genes and viral genes in insect culture cells. The BiFC assay confirmed the specificity of the interactions including B. mori 35 kDa protease and two BmDNV-Z proteins encoded by VD1-ORF2 and VD2-ORF1; B. mori transgelin and BmDNV-Z protein encoded by VD2-ORF3; and B. mori serine protease precursor and BmDNV-Z encoded by VD2-ORF3 in vitro. Our findings revealed that the specific host midgut proteins are involved in the interactions between B. mori and BmDNV-Z, which will facilitate our understanding of the molecular mechanisms of BmDNV-Z infection. @@@ Structured digital abstract BmDNV VD1-ORF4 physically interacts with trypsin-like protease by two hybrid pooling approach (View interaction) 35 kDa protease and BmDNV VD2-ORF1 physically interact by bimolecular fluorescence complementation (View interaction) 35 kDa protease and BmDNV VD1-ORF2 physically interact by bimolecular fluorescence complementation (View interaction) BmDNV VD1-ORF4 physically interacts with alpha amylase by two hybrid pooling approach (View interaction) trypsin-like protease physically interacts with BmDNV VD1-ORF4 by two hybrid (View interaction) BmDNV VD1-ORF4 physically interacts with translation elongation factor 2 by two hybrid pooling approach (View interaction) BmDNV VD2-ORF1 physically interacts with ribosomal protein S5 by two hybrid pooling approach (View interaction) translation elongation factor 2 physically interacts with BmDNV VD1-ORF4 by two hybrid (View interaction) BmDNV VD1-ORF2 physically interacts with olfactory receptor by two hybrid pooling approach (View interaction) olfactory receptor physically interacts with BmDNV VD1-ORF2 by two hybrid (View interaction) BmDNV VD1-ORF2 physically interacts with 35 kDa protease by two hybrid pooling approach (View interaction) BmDNV VD1-ORF4 physically interacts with olfactory receptor by two hybrid pooling approach (View interaction) BmDNV VD2-ORF1 physically interacts with 35 kDa protease by two hybrid pooling approach (View interaction) BmDNV VD2-ORF3 physically interacts with serine protease precursor by two hybrid pooling approach (View interaction) 35 kDa protease physically interacts with BmDNV VD1-ORF2 by two hybrid (View interaction) BmDNV VD1-ORF1 physically interacts with ribosomal protein S5 by two hybrid pooling approach (View interaction) BmNPV LEF-9 and BmNPV P47 physically interact by bimolecular fluorescence complementation (View interaction) BmDNV VD1-ORF4 physically interacts with chymotrypsin-like protein by two hybrid pooling approach (View interaction) transgelin physically interacts with BmDNV VD2-ORF3 by two hybrid (View interaction) serine protease precursor and BmDNV VD2-ORF3 physically interact by bimolecular fluorescence complementation (View interaction) transgelin and BmDNV VD2-ORF3 physically interact by bimolecular fluorescence complementation (View interaction) 30 kP protease A physically interacts with BmDNV VD1-ORF4 by two hybrid (View interaction) serine protease precursor physically interacts with BmDNV VD2-ORF3 by two hybrid (View interaction) BmDNV VD2-ORF3 physically interacts with transgelin by two hybrid pooling approach (View interaction) BmDNV VD1-ORF4 physically interacts with 35 kDa protease by two hybrid pooling approach (View interaction) BmDNV VD1-ORF4 physically interacts with 30 kP protease A by two hybrid pooling approach (View interaction) ribosomal protein S5 physically interacts with BmDNV VD1-ORF1 by two hybrid (View interaction) carboxypeptidase physically interacts with BmDNV VD1-ORF4 by two hybrid (View interaction) BmDNV VD1-ORF4 physically interacts with carboxypeptidase by two hybrid pooling approach (View interaction)
- 出版日期2013-2
- 单位浙江大学
