家蚕中猪细小病毒病毒样颗粒的制备OA北大核心

In order to prevent and control porcine parvovirus(PPV)infection,this study utilized silkworm baculovirus expression system to prepare PPV virus-like particles(VLPs).Firstly,the VP2 gene of PPV structural protein was amplified and cloned into the pFastBac Ⅰ vector,constructing the transfer vector pFastBac Ⅰ-VP2.The transfer vector pFastBac Ⅰ-VP2 was transformed into Escherichia coli to prepare recombinant Bacmid.The recombinant Bacmid was transfected into silkworm ovary(BmN)cells to prepare recombinant baculovirus.The expression of VP2 protein was identified by Western blot,the morphology of PPV VLPs was observed using transmission electron microscopy,and the titer of PPV VLPs was determined by hemagglutination assay.PPV VLPs were prepared by injecting silkworm larvae with recombinant baculovirus containing the PPV VP2 gene,and the PPV VLPs in silkworms were identified.The results showed that in both BmN cell samples and silkworm larval samples,a target band of VP2 protein was observed at around 64 kDa in Western blot.PPV VLPs with a diameter of about 23 nm were observed under transmission electron microscopy.The hemagglutination titer of PPV VLPs prepared in BmN cells was 1:26,and in silkworms,it was 1:29,higher than that prepared in BmN cells.The results indicate that PPV VLPs were successfully prepared in silkworms in this study,and they exhibited good biological activity,providing data support for the development of PPV VLPs vaccines.