抗番茄花叶病毒RNAi载体的构建及烟草遗传转化  

Construction of RNAi Vector Which Resist Tomato mosaic virus and Transformation of Tobacco

作　　者：赵峰玉[1,2] 郑银英[1,2] 张瑜[1,2] 乔亚红[1,2] 崔百明[1,2] 向本春[1,2] 

机构地区：[1]石河子大学农业生物技术重点实验室 [2]石河子大学生命科学院,石河子832003

出　　处：《石河子大学学报（自然科学版）》2011年第2期163-168,共6页Journal of Shihezi University(Natural Science)

基　　金：国际科技合作项目(2008DFA30560);973计划前期研究专项(2008CB117018)

摘　　要：为了获得抗番茄花叶病毒的转基因烟草,试验采用RT-PCR技术扩增新疆加工番茄花叶病毒基因组,然后与已知株系序列比对获得相应的保守序列。根据保守区选择干扰序列并设计引物进行PCR扩增,构建了pBi35STo5,pBi35SToBK,pBi35ToMV3 3个RNAi表达载体;利用农杆菌介导法分别将表达载体侵染普通烟,转化植株经分子鉴定,结果显示已成功获得了转基因阳性植株,为进一步的攻毒试验奠定了基础。To obtain transgenic tobacoo which resist Tomato mosaic virus,genome sequence of ToMV from Xinjiang processing tomato was amplified by reverse transcription polymerase chain reaction（RT-PCR）,then the conserved sequence of the virus among known strains were obtained by using sequence alignment;RNAi sequences were chosen according to conserved regions,and amplified by PCR,pBi35STo5,pBi35SToBK,pBi35ToMV3 three RNAi expression vectors were constructed;through introducing vectors into Nicotiana occidentalis by Agrobacterium mediated transformation separately.The results showed that positive transgenic plants were obtained successfully by molecular identification,which also provides a foundation for further experiment of attacking virus.

关 键 词：加工番茄 RNAI 番茄花叶病毒 

分 类 号：S436.412.19[农业科学—农业昆虫与害虫防治]