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当前位置: 首页 > 文章 > 应用TALENs技术定向敲除烟草eIF4E-6基因 安徽农业科学 2017,45 (4) 142-146
Position: Home > Articles > The Directional Knockout of eIF4E-6 Gene Using TALENs Technique Journal of Anhui Agricultural Sciences 2017,45 (4) 142-146

应用TALENs技术定向敲除烟草eIF4E-6基因

作  者：
宋琳娜;杨鹏九;万秀清;乔婵;潘洪杏;郭振楠;刘侠;郭兆奎;李若
单  位：
牡丹江医学院;牡丹江师范学院;牡丹江烟草科学研究所;黑龙江省烟草专卖局
关键词：
烟草;PVY;K326;eIF4E-6基因;TALENs
摘  要：
[目的]利用TALENs技术定向敲除烟草eIF4E-6基因,为提高优质烤烟品种K326的马铃薯Y病毒(PVY)抗性提供材料。[方法]构建特异靶向烟草eIF4E-基因的TALENs载体,并由农杆菌介导转化K326烟草,用PCR和克隆测序方法筛选目的基因被成功编辑的T_0代转基因烟株。[结果]构建了2个特异靶向eIF4E-6基因的TALENs载体T_3和T_4。2个载体各获得了约40和50株阳性转化的K326 T_0代烟株。T_3载体获得1株目的基因靶位点发生大片段碱基缺失的杂合突变型T_0代单株、3株靶位点发生碱基替换的杂合突变型T_0代单株。T_4载体获得5株靶位点发生碱基替换的杂合突变型T_0代单株。[结论]利用TALENs技术获得的eIF4E-6基因杂合突变型K326 T_0代单株为后续获得纯合基因敲除的K326烟草,并最终获得PVY病毒抗性改良的K326烟草材料奠定了基础。
译  名：
The Directional Knockout of eIF4E-6 Gene Using TALENs Technique
作  者：
SONG Lin-na;YANG Peng-jiu;WAN Xiu-qing;LI Ruo;Mudanjiang Normal University;Mudanjiang Tobacco Science Research Institute;Heilongjiang Tobacco Monopoly Bureau;
关键词：
Tobacco;;PVY;;K326;;eIF4E-6 gene;;TALENs
摘  要：
[Objective] To knockout eIF4E-6 gene using TALENs technique,and to provide material for improving the PVY resistance of highclass flue-cured tobacco cultivar K326.[Method]TALENs vectors that specifically targeting against eIF4E-6 gene were constructed. TALENs vectors transformed tobacco cultivar K326 mediated by Agrobacterium tumefaciens. The T_0 transgenic tobacco plants of which eIF4E-6 gene were successfully edited were screened.[Result] Two TALENs vectors which specifically targeted against eIF4E-6 gene were constructed in this study.Two vectors obtained approximately 40 and 50 positively transformed T_0 tobacco plants respectively. Vector T_3 obtained 1 heterozygous mutant T_0 plant whose targeting site of eIF4E-6 gene showed deletion of big base fragment and 3 heterozygous mutant plants whose targeting sites showed single base substitution. Vector T_4 obtained 5 heterozygous mutant T_0 plants whose targeting sites showed single base substitution. [Conclusion]These heterozygous mutant T_0K326 tobacco plants obtained by using TALENs technique laid foundation for obtaining homozygous gene knockout K326 tobacco and finally obtaining K326 tobacco with improved PVY resistance.

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