应用Gateway^(TM)技术构建烟草抗真菌病相关基因的RNAi载体  

Construction of Tobacco RNAi Vectors for Fungi Resistance-related Genes Using Gateway^(TM) Technology

作　　者：赵百英[1] 李凤霞[1] 杨爱国[1] 孙玉合[1] 苏振刚 杨帆[1] 刘慧[1] 王元英[1] 

机构地区：[1]中国农业科学院烟草研究所/烟草遗传改良与生物技术重点开放实验室,山东青岛266101 [2]红塔烟草(集团)有限责任公司,云南玉溪653100

出　　处：《湖北农业科学》2013年第5期1190-1195,共6页Hubei Agricultural Sciences

基　　金：国家烟草专卖局重点项目(110200701022;110201002002)

摘　　要：根据抗病基因(Resistance genes,R基因)的功能保守域,利用生物信息分析方法在绒毛状烟草(Nicotiana tomentosiformis)全基因组上获得6个完整可靠的抗真菌病候选基因。在这6个基因的非保守区设计特异引物,以普通烟草品种红花大金元反转录cDNA为模板克隆抗真菌病相关基因的干扰片段,利用GatewayTM克隆技术构建上述候选基因的RNAi载体,经酶切和测序证明载体构建正确,为进一步鉴定这6个候选基因的具体抗病性,并为将其应用于烟草抗病新品种的选育工作奠定了基础。According to the conserved domain of resistance genes(R gene),six disease resistance candidate genes were obtained from the genome of Nicotiana tomentosiformis by bioinformatics.Specific primers was designed according to the non-conservative regions of these six genes,and applied for cloning of interference clips of fungus-resistant genes using reverse transcriptase cDNA of N.tobacum cv.Honghuadajinyuan as templates.Then the RNAi vectors of these six candidate genes were constructed through GatewayTM cloning techniques.According to restriction endonuclease digestion and sequencing,the RNAi vectors was correctly constructed,which lay a foundation for further identification of the six candidate genes and applying in breeding of tobacco disease resistant varieties.

关 键 词：烟草(Nicotiana) GatewayTM技术 RNAI载体 R基因 相似性 结构域 

分 类 号：S[农业科学]