口蹄疫病毒L、2B基因siRNA重组腺病毒质粒的构建  

Construction of Recombinant Adenovirus Plasmid Containing the L and 2B Gene si RNAs of Foot-and-Mouth Disease Virus

作　　者：贾俊涛[1] 郭凤英[1] 乌伦吉如嘎[1] 张小宇[1] 陈金顶[2] 

机构地区：[1]内蒙古农业大学职业技术学院,内蒙古包头014109 [2]华南农业大学,广州510642

出　　处：《湖北农业科学》2017年第16期3152-3154,共3页Hubei Agricultural Sciences

基　　金：教育部"长江学者和创新团队发展计划"创新团队项目(TRT0723);广东省自然科学基金创新团队项目(5200638);广东省自然科学基金项目(020995;06025827);广州市科技计划项目(2008Z1-E011)

摘　　要：根据si RNA设计原则,分别选择O型FMDV的L和2B基因上保守序列作为可能的干扰位点,设计了2个si RNAs,并将si RNAs克隆到p SIREN-Shuttle中,获得2个si RNA重组表达载体p Shuttle-L和p Shuttle-2B。用限制性内切酶I-Ceu I和PI-Sce I双酶切si RNA重组表达载体和腺病毒质粒载体p Adeno-X,利用体外连接法获得了重组质粒。经PCR扩增、酶切鉴定及序列测定,成功构建了重组腺病毒质粒p Adeno-L和p Adeno-2B。In this study, the conservative sequence in L and 2B gene of O type FMDV were chose as possible interference sites. Two siRNAs were designed and synthesized. These siRNAs were cloned into the plasmid pSIREN-Shuttle, then the siR- NAs recombined expression vectors were obtained including pShuttle-L and pShuttle-2B. The vectors and the adenovirus vec- tor pAdeno-X were digested with restriction endonuclease I-Ceu I and PI-Sce I and connected in vitro, then recombinant plasmids were obtained. The result of PCR, enzyme-cutting and sequencing suggested that the recombined adenovirus plas- raids pAdeno-L and pAdeno-2B were constructed successfully.

关 键 词：口蹄疫病毒 RNA干扰 重组腺病毒质粒 

分 类 号：S852.659.6[农业科学—基础兽医学]