title: Baculovirus-mediated GCRV vp7 and vp6 genes expression in silkworm and grass carp.
abstract: Grass carp hemorrhagic disease is a common fish disease and often results in significant economic losses in grass carp aquaculture in China. This study was aimed to develop a novel oral vaccine against grass carp reovirus (GCRV). GCRV vp6 and vp7 genes with β-actin promoter of Megalobrama amblycephala and polyhedrin promoter (Ph10) of baculovirus, respectively, were cloned into plasmid pFast™-Dual to construct a vector pFast-PHVP7-AVP6, which was used to generate a recombinant baculovirus BacFish-vp6/vp7 via Bac-to-Bac system. The VP7 expression was analyzed from freeze-dried powder of the BacFish-vp6/vp7-infected silkworm pupae by western blotting, and VP6 expression was analyzed from orally vaccinated fish with the freeze-dried powder by RT-PCR. The VP6 expression was also analyzed from both CIK cells transduced with BacFish-vp6/vp7 and tissues of vaccinated fish by immunofluorescence analysis. Recombinant VP7 could be detected from the BacFish-vp6/vp7-infected silkworm pupae. Pathological changes were not observed in CIK cells transduced with BacFish-vp6/vp7, and VP6 expression was found in CIK cells. When the grass carps were orally administrated with the freeze-dried powder, vp6 gene transcription was found in blood of the vaccinated fishes and VP6 protein was observed in liver and kidney of the vaccinated fish by immunofluorescence analysis. These results indicated that vp7 gene was expressed in the BacFish-vp6/vp7-infected silkworm and vp6 gene was expressed in orally vaccinated fish with freeze-dried powder of the BacFish-vp6/vp7-infected silkworm pupae, suggesting the possibility to use the powder as an orally administrated vaccine.
authors.affiliation: School of Biology & Basic Medical Science, Soochow University, No. 199 Ren-ai Road, Suzhou, 215123, Jiangsu, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Liu
authors.affiliation: School of Biology & Basic Medical Science, Soochow University, No. 199 Ren-ai Road, Suzhou, 215123, Jiangsu, China.
authors.forename: Yongchang
authors.initials: Y
authors.lastname: Gong
authors.affiliation: School of Biology & Basic Medical Science, Soochow University, No. 199 Ren-ai Road, Suzhou, 215123, Jiangsu, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Li
authors.affiliation: School of Biology & Basic Medical Science, Soochow University, No. 199 Ren-ai Road, Suzhou, 215123, Jiangsu, China.
authors.forename: Xiaolong
authors.initials: X
authors.lastname: Hu
authors.affiliation: School of Biology & Basic Medical Science, Soochow University, No. 199 Ren-ai Road, Suzhou, 215123, Jiangsu, China.
authors.forename: Guangli
authors.initials: G
authors.lastname: Cao
authors.affiliation: School of Biology & Basic Medical Science, Soochow University, No. 199 Ren-ai Road, Suzhou, 215123, Jiangsu, China.
authors.forename: Renyu
authors.initials: R
authors.lastname: Xue
authors.affiliation: School of Biology & Basic Medical Science, Soochow University, No. 199 Ren-ai Road, Suzhou, 215123, Jiangsu, China. gongcl@suda.edu.cn.
authors.forename: Chengliang
authors.initials: C
authors.lastname: Gong
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D012831:Bombyx / Q000235:genetics; D002347:Carps / Q000821:virology*; D002460:Cell Line; D003001:Cloning, Molecular; D005393:Fish Diseases / Q000517:prevention & control / Q000821:virology*; D005398:Fisheries; D015870:Gene Expression; D012333:RNA, Messenger / Q000097:blood / Q000235:genetics; D012367:RNA, Viral / Q000097:blood; D012087:Reoviridae / Q000235:genetics*; D012088:Reoviridae Infections / Q000517:prevention & control / Q000662:veterinary* / Q000821:virology; D014611:Vaccination; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis / Q000235:genetics*; D014765:Viral Vaccines / Q000235:genetics / Q000276:immunology
chemical_list: D012333:RNA, Messenger; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines
keywords: Baculovirus expression system; Bombyx mori; Grass carp reovirus (GCRV); Oral vaccine; vp6 gene; vp7 gene