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本氏烟抗细胞凋亡基因NbDAD1克隆及遗传转化研究 被引量：1

Cloning and Geneic Transformation of Nbdad1 Gene from Nicotiana benthamiana
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摘要 [目的]克隆本氏烟的抗细胞凋亡基因NbDAD1,并对该基因进行遗传转化研究。[方法]通过RT-PCR技术分离本氏烟NbDAD1基因,构建基因超表达载体,以获得转入NbDAD1基因和携带HAtag标签的NbDAD1基因抗性植株。[结果]试验克隆出了全长351 bp的本氏烟NbDAD1基因;成功构建了重组质粒pCAMBIA1301-NbDAD1和pCAMBIA1301-NbDAD1HAtag;共获得3个基因型50株T0代抗Hyg本氏烟植株,其中23株检测PCR呈阳性。[结论]该试验为研究NbDAD1基因在本氏烟中的具体功能及深入探讨DAD1蛋白在植物细胞程序性死亡中可能的作用机制奠定了基础。 [Objective] This study aimed to clone NbDAD1 gene from N.benthamiana and investigate its genetic transformation.[Method] NbDAD1 gene was isolated from N.benthamiana by using RT-PCR technology and the over-expression vectors were constructed to obtain Nbdad1-overexpression resistant plants and Nbdad1-overexpression resistant plants carrying HA tag.[Result] The 351 bp NbDAD1 gene was cloned from N.benthamiana;recombinant plasmids pCAMBIA1301-NbDAD1 and pCAMBIA1301-NbDAD1HAtag were constructed successfully;50 T0-generation N.benthamiana Hyg-resistant transgenic lines of three genotypes were obtained,including 23 positive transgenic plants.[Conclusion] This study laid the foundation for investigating the specific functions of NbDAD1 gene in N.benthamiana and exploring the possible functional mechanism of DAD1 protein in programmed cell death of plants.
作者 徐玲玲 陈崇崇 王梨嬛 潘永娟 杨莉
机构地区 浙江师范大学化学与生命科学学院
出处 《安徽农业科学》 CAS 2012年第16期8797-8799,8805,共4页 Journal of Anhui Agricultural Sciences
基金 浙江省自然科学基金项目(Y3110409)
关键词 NbDAD1基因 遗传转化 本氏烟 PCD NbDAD1 gene Genetic transformation Nicotiana benthamiana PCD
分类号 S188.1 [农业科学—农业基础科学]
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参考文献23

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共引文献77

同被引文献50

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安徽农业科学
2012年 第16期

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