以减毒鼠伤寒沙门氏菌为载体的重组NDV-HN口服疫苗的构建与鉴定  

Construction and Identification of Recombinant Attenuated Salmonella typhimurium Oral NDV-HN Vaccine

作　　者：田芳华[1] 程相朝[1] 张春杰[1] 李银聚[1] 李静[1] 刘一尘[1] 

机构地区：[1]河南科技大学动物科技学院,河南洛阳471003

出　　处：《中国畜牧兽医》2013年第11期59-63,共5页China Animal Husbandry & Veterinary Medicine

基　　金：河南科技大学研究生创新基金(CXJJ-YJS-Z005);河南省科技公关项目(112102110019)

摘　　要：试验旨在构建表达鸡新城疫病毒(NDV)HN基因的重组减毒鼠伤寒沙门氏菌SL1344ΔcrpΔasd(pYA3493-HN)。以pMD18-T-HN为模板,通过PCR扩增出NDV HN基因片段,定向插入原核表达载体pYA3493中,将重组表达质粒pYA3493-HN转入χ6097,再转入减毒鼠伤寒沙门氏菌SL1344ΔcrpΔasd,通过双酶切和PCR对质粒进行鉴定。结果表明,携带NDV HN基因片段的重组减毒鼠伤寒沙门氏菌SL1344ΔcrpΔasd(pYA3493-HN)构建成功。本研究结果为开发鸡新城疫的口服基因工程活载体疫苗奠定了基础。The study was aimed to construct prokaryotic expression vector of NDV HN gene using attenuated Salmonella t：yphimurium SL1344AcrpAasd as a carrier. The HN gene was cloned from the plasmid pMD18-T-HN by polymerase chain re- action （PCR） and inserted into the vector pYA3493. Electricity transformation method was used in this recombinant plasmid pYA3493-HN transformating to recepted state Salmonella typhimurium SL1344AcrpAasd. The recombinant plasmid pYA3493-HN was identified by digestion of endonuclease, PCR and sequencing. The results confirmed that recombinant atten- uated Salmonella typhimurium vaccine SL1344AcrpAasd （pYA3493-HN） was constructed successfully, which provided a foundation for research and development of NDV-HN orally genqtic engineering live vaccine.

关 键 词：新城疫 HN基因 原核表达载体pYA3493 减毒鼠伤寒沙门氏菌 

分 类 号：S852.612[农业科学—基础兽医学]