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.
出处
《中国畜牧兽医》
CAS
北大核心
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)
