2016
DOI: 10.1371/journal.pone.0152140
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A Highly Efficient and Simple Construction Strategy for Producing Recombinant Baculovirus Bombyx mori Nucleopolyhedrovirus

Abstract: The silkworm baculovirus expression system is widely used to produce recombinant proteins. Several strategies for constructing recombinant viruses that contain foreign genes have been reported. Here, we developed a novel defective-rescue BmNPV Bacmid (reBmBac) expression system. A CopyControl origin of replication was introduced into the viral genome to facilitate its genetic manipulation in Escherichia coli and to ensure the preparation of large amounts of high quality reBmBac DNA as well as high quality reco… Show more

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Cited by 10 publications
(24 citation statements)
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“…Human serum was obtained from Sigma-Aldrich (Deisenhofen, Germany). The reBmBac vector was constructed and prepared in our lab [ 42 ]. Expression of EGFP was analyzed by western blotting using the rabbit polyclonal anti-EGFP antibody (ab6556, Abcam, UK).…”
Section: Methodsmentioning
confidence: 99%
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“…Human serum was obtained from Sigma-Aldrich (Deisenhofen, Germany). The reBmBac vector was constructed and prepared in our lab [ 42 ]. Expression of EGFP was analyzed by western blotting using the rabbit polyclonal anti-EGFP antibody (ab6556, Abcam, UK).…”
Section: Methodsmentioning
confidence: 99%
“…The recombinant Bm NPVs for reporter gene delivery, reBm-luc and reBm-EGFP, were prepared by co-transfection in Bm 5 cells with reBmBac and pVLCMV-luc or pVLCMV-EGFP as described [ 42 , 43 ], and purified by plaque screening [ 44 ]. Fifth instar silkworm larvae were used to amplify the viruses; they were injected with recombinant Bm NPVs (approximately 10 5 plaque forming units, p.f.u.).…”
Section: Methodsmentioning
confidence: 99%
“…We have previously demonstrated the successful construction and development of a recombinant B. mori Bacmid (reBmBac) expression system [41]. The silkworm reBmBac gene expression system was used to construct and express recombinant chIFN-γ (GenBank: NM_205149.1; Gallus gallus).…”
Section: Expression Of Chifn-γmentioning
confidence: 99%
“…Blank silkworm blood treated in the same way was loaded as the negative control. After electrophoresis and transfer onto PVDF membrane, recombinant chIFN-γ expression was analyzed by western blot analysis using anti-IFN-γ antibody (Abcam, Cambridge, USA), according to the Protein Blotting Guide (Bio-Rad, Hercules, USA), as previously described [41].…”
Section: Western Blot Analysismentioning
confidence: 99%
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