| 本氏烟抗细胞凋亡基因NbDAD1克隆及遗传转化研究 |
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| 引用本文： | 徐玲玲,陈崇崇,王梨嬛,潘永娟,杨莉. 本氏烟抗细胞凋亡基因NbDAD1克隆及遗传转化研究[J]. 安徽农业科学, 2012, 40(16): 8797-8799,8805 |
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| 作者姓名： | 徐玲玲 陈崇崇 王梨嬛 潘永娟 杨莉 |
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| 作者单位： | 浙江师范大学化学与生命科学学院,浙江金华,321004;浙江师范大学化学与生命科学学院,浙江金华,321004;浙江师范大学化学与生命科学学院,浙江金华,321004;浙江师范大学化学与生命科学学院,浙江金华,321004;浙江师范大学化学与生命科学学院,浙江金华,321004 |
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| 基金项目： | 浙江省自然科学基金项目(Y3110409) |
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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。
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| 关 键 词： | NbDAD1基因 遗传转化 本氏烟 PCD |
Cloning and Geneic Transformation of Nbdad1 Gene from Nicotiana benthamiana |
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| Affiliation: | XU Ling-ling et al(College of Chemistry and Life Science,Zhejiang Normal University,Jinhua,Zhejiang 321004) |
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| Abstract: | [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. |
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| Keywords: | NbDAD1 gene Genetic transformation Nicotiana benthamiana PCD |
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