[Objective] Sheep contagious ecthyma virus B2L gene recombinant adenovirus was built by adenovirus vector system.[Method] Genome DNA extracted from sheep contagious ecthyma virus strain JLSY04 as a template,Gene fragm...[Objective] Sheep contagious ecthyma virus B2L gene recombinant adenovirus was built by adenovirus vector system.[Method] Genome DNA extracted from sheep contagious ecthyma virus strain JLSY04 as a template,Gene fragments obtained from B2L by PCR amplification;B2L gene cloning was cloned into PDNR-CMD vector,screening positive clones and plasmid CTC572-6 was obtained;CTC572-6 plasmid for homologous was recombined with the adenoviral vector.Screening positive clones and bacilli PCR,digestion and sequencing and so on were identified.[Result] After identified by enzyme digestion and gene sequencing,recombinant adenovirus vector CTC572Ade-30 of carrying sheep contagious ecthyma virus B2L gene was constructed successfully.[Conclusion] Which laid the foundation for sheep contagious ecthyma genetically engineered vaccine.展开更多
本研究利用北京甘蓝CMV分离物(CMV-BG)全长2b基因,构建了含其正义和反义序列的PVX侵染性载体pVX2bS和pVX2bAS,并进行了它们与PVX侵染性载体间的共侵染稳定性实验。结果表明:pVX2bS和pVX2bAS能侵染本氏烟、SR1、Sam sun NN烟草、Shepody...本研究利用北京甘蓝CMV分离物(CMV-BG)全长2b基因,构建了含其正义和反义序列的PVX侵染性载体pVX2bS和pVX2bAS,并进行了它们与PVX侵染性载体间的共侵染稳定性实验。结果表明:pVX2bS和pVX2bAS能侵染本氏烟、SR1、Sam sun NN烟草、Shepody马铃薯和中蔬5号番茄,与pSfinx相比,表现不同程度的症状延迟效应;pVX2bS和pVX2bAS在22℃/16℃(昼/夜温度)条件下,接种SR1烟草45d仍可检测到重组载体的存在;pVX2bS和pVX2bAS菌液混合接种15d后只能检测到正义或反义一种载体的随机表达,二者交互接种,后接种的一方不能表达。展开更多
基金Support by Science and Technology Development Program of Jilin Province(20080106)~~
文摘[Objective] Sheep contagious ecthyma virus B2L gene recombinant adenovirus was built by adenovirus vector system.[Method] Genome DNA extracted from sheep contagious ecthyma virus strain JLSY04 as a template,Gene fragments obtained from B2L by PCR amplification;B2L gene cloning was cloned into PDNR-CMD vector,screening positive clones and plasmid CTC572-6 was obtained;CTC572-6 plasmid for homologous was recombined with the adenoviral vector.Screening positive clones and bacilli PCR,digestion and sequencing and so on were identified.[Result] After identified by enzyme digestion and gene sequencing,recombinant adenovirus vector CTC572Ade-30 of carrying sheep contagious ecthyma virus B2L gene was constructed successfully.[Conclusion] Which laid the foundation for sheep contagious ecthyma genetically engineered vaccine.
文摘本研究利用北京甘蓝CMV分离物(CMV-BG)全长2b基因,构建了含其正义和反义序列的PVX侵染性载体pVX2bS和pVX2bAS,并进行了它们与PVX侵染性载体间的共侵染稳定性实验。结果表明:pVX2bS和pVX2bAS能侵染本氏烟、SR1、Sam sun NN烟草、Shepody马铃薯和中蔬5号番茄,与pSfinx相比,表现不同程度的症状延迟效应;pVX2bS和pVX2bAS在22℃/16℃(昼/夜温度)条件下,接种SR1烟草45d仍可检测到重组载体的存在;pVX2bS和pVX2bAS菌液混合接种15d后只能检测到正义或反义一种载体的随机表达,二者交互接种,后接种的一方不能表达。