Development of recombinant vaccines against canine protozoan diseases by using canine herpes virus vector

1.犬疱疹病毒载体的建立本研究对犬疱疹病毒的毒力基因、非必需基因如TK、gE和gI基因进行了鉴定和克隆。缺失这些基因的犬疱疹病毒可以在体外生长，但其在体内的毒力大大降低。2.犬寄生虫免疫优势抗原基因的克隆对犬致病性寄生虫吉氏巴氏巴氏杆菌和犬新孢子虫的免疫优势抗原基因进行了鉴定、克隆和鉴定。主要基因区为：B.gibosoni BgP 29、BgP 50、BgP 76、BgTRAP、BgAMA-1。BgRAP-1、Bg12D3 ;表达原虫抗原的重组病毒的构建构建表达BgP 29、BgP 50、BgP 76、NcSAG 1、NcSRS 2、NcGRA 7、NcPDI、NcRP 1、NcHSP 70的重组犬疱疹病毒或重组牛痘病毒，并证明重组病毒表达的重组抗原与寄生虫的天然抗原相似，4.重组疫苗的免疫保护效果用上述重组病毒免疫的犬分别产生了抗吉氏B.gibsoni和犬奈瑟氏菌感染的部分保护性免疫。

1.Development of canine herpesvirus vectorIn the present study, some virulent and non-essential genes, such as TK, gE and gI genes of canine herpesviruse were identified and cloned. Canine herpesviruse deleting these genes can grow in vitro, however their virulence reduced greatly in vivo. Therefore, these genes were proposed as targets for insertion of foreign immunogenic genes.2.Cloning of genes encoding immunodominant antigens of canine protozoan parasitesMany genes encoding immunodominat antigens of canine pathogenic protozoan parasites, Babesia gibsoni and Neospora caninum were identified, cloned and characterized. The main genes areas follows : B.gibosoni BgP29, BgP50, BgP76, BgTRAP, BgAMA-1. BgRAP-1, Bg12D3 ; N.caninum NcSAG1, NcSRS2, NcGRA7, NcPDI, NcRP1, NcHSP70.3.Construction of recombinant viruses expressing protozoan antigensRecombinant canine herpesvirus or vaccinia virus expressing BgP29, BgP50, BgP76, NcSAG1, NcSRS2 were constructed, and demonstrated that the recombinant antigens expressed by recombinant viruses were similar to their native antigens from parasites in antigenicity.4.Protective effects of recombinant vaccinesDogs immunized with above recombinant viruses produced partial protective immunity against B.gibsoni and N.caninum infections, respectively.