运送鹅源新城疫病毒DNA疫苗的减毒沙门氏菌及其免疫原性  

DNA Vaccine Against Newcastle Disease Virus of Goose Origin Delivered by Attenuated Salmonella typhimurium and Its Immunogenicity for Mice

作　　者：潘志明[1] 焦新安[1,2] 张小荣[3] 张晓明[1] 黄金林[1] 万洪全[3] 刘秀梵[3] 

机构地区：[1]扬州大学生物科学与技术学院 [2]扬州大学农业部畜禽传染病学重点开放实验室扬州225009 [3]扬州大学农业部畜禽传染病学重点开放实验室

出　　处：《农业生物技术学报》2004年第4期412-415,共4页Journal of Agricultural Biotechnology

基　　金：国家高技术研究与发展计划(863)项目(No.2002AA245051);国家自然科学基金项目(No.30170700);江苏省自然科学基金项目(No. BK2003043)。

摘　　要：根据GenBank中发表的新城疫病毒融合蛋白(F)基因序列,设计一对引物,通过RT-PCR扩增出鹅源新城疫病毒分离株JS5 F基因,约1 700 bp,测序确认后,将其克隆入真核表达载体pVAX1,得到候选DNA疫苗pVAX1-F。将pVAX1-F转化减毒鼠伤寒沙门氏菌(Salmonellatyphimurium)X4550,获得运送DNA疫苗的重组沙门氏菌X4550(pVAX1-F)。以2×108CFU/只的剂量两次免疫CD1小鼠,免疫小鼠可以检测到特异性针对NDV F蛋白的血清抗体应答,X4550(pVAX1-F)免疫组抗体水平显著高于X4550(pVAX1)组(P <0.05),表明该运送DNA疫苗的减毒沙门氏菌系统在体内能够成功释放所携带的质粒,并能够刺激机体产生免疫应答。A pair of primers were designed and synthesized according to the previously published sequence of fusion protein (F) gene of Newcastle disease virus (NDV) and used to amplify F gene by reverse-transcription polymerase chain reaction (RT-PCR) from the genomic RNA of a NDV strain JS5 isolated from goose. The PCR product was identified by sequencing. Then recombinant eukaryotic expression vector pVAX1-F was constructed through inserting F gene into MCS of pVAX1. Finally, the recombinant vector was transformed into attenuated Salmonella typhimurium X4550, the recombinants were screened and the positive clone was designated as X4550 (pVAX1-F). This recombinant bacteria could significantly elicit specific antibody response in immunized CD1 mice compared with X4550(pVAX1) immunized mice. These results demonstrated that the Salmonella delivering system could release haboured NDV DNA vaccine in vivo and elicit spcific immune response.

关 键 词：DNA疫苗 减毒沙门氏菌 鹅源新城疫病毒 免疫原性 运送 减毒鼠伤寒沙门氏菌 GENBANK 真核表达载体 病毒分离株 PCR扩增 基因序列 融合蛋白 抗体应答 免疫小鼠 免疫应答 F基因 F蛋白 NDV 特异性 

分 类 号：S942.5[农业科学—水产养殖] S858.315.3[农业科学—水产科学]