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羊传染性脓疱病毒B2L基因片段重组腺病毒载体的构建
引用本文:邵洪泽,毛文智,宋阳,李琳,程荣华.羊传染性脓疱病毒B2L基因片段重组腺病毒载体的构建[J].安徽农业科学,2010,38(13):6722-6724.
作者姓名:邵洪泽  毛文智  宋阳  李琳  程荣华
作者单位:吉林省畜牧兽医科学研究院,吉林长春,130062;吉林农业大学,吉林长春,130118;吉林省畜牧兽医科学研究院,吉林长春,130062;吉林省食品工业产品质量监督监测站,吉林长春,130061
基金项目:吉林省科技发展计划项目 
摘    要:目的]利用腺病毒载体系统构建羊传染性脓疱病毒B2L基因重组腺病毒载体。方法]以从羊传染性脓疱病毒株JLSY04中提取的基因组DNA为模板,PCR扩增获得B2L目的基因片段;然后将B2L目的基因克隆至PDNR-CMV载体,筛选阳性克隆获得质粒CTC572-6;再将质粒CTC572-6与腺病毒载体进行同源重组,筛选阳性克隆,并进行菌液PCR、酶切、测序等鉴定。结果]经酶切和基因测序等鉴定,成功构建了携带羊传染性脓疱病毒B2L基因的重组腺病毒载体CTC572Ade-30。结论]为羊传染性脓疱基因工程疫苗的进一步研究奠定基础。

关 键 词:羊传染性脓疱病毒  B2L基因  腺病毒载体

Construction of Recombinant Adenovirus Vector Containing CEVB2L Gene
Institution:SHAO Hong-ze et al (Jilin Academy of Animal Science and Veterinary Medicine,Changchun,Jilin 130062)
Abstract:Objective] Sheep contagious pustular dermatitis virus B2L recombinant adenovirus was built by adenovirus vector system. Method] Genome DNA extracted from sheep contagious pustular dermatitis virus strain as a template,gene fragments obtained from B2L by PCR amplification; B2L gene 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 pustular dermatitis virus B2L gene was constructed successfully. Conclusion] The study laid the foundation for sheep contagious pustular dermatitis genetically engineered vaccine.
Keywords:CEV  B2L gene  Adenovirus vector
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