CAJ | 학술논문

[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351bp的本氏烟NbDAD1基因；成功构建了重组质粒pCAMBIA1301．NbDADI和pCAMBIA1301．NbDAD1HAtag；共获得3个基因型50株R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。
[목적]극륭본씨연적항세포조망기인NbDAD1，병대해기인진행유전전화연구。[방법]통과RT—PCR기술분리본씨연NbDAD1기인，구건기인초표체재체，이획득전입NbDAD1기인화휴대HAtag표첨적NbDAD1기인항성식주。[결과]시험극륭출료전장351bp적본씨연NbDAD1기인；성공구건료중조질립pCAMBIA1301．NbDADI화pCAMBIA1301．NbDAD1HAtag；공획득3개기인형50주R대항Hyg본씨연식주，기중23주검측PCR정양성。[결론]해시험위연구NbDAD1기인재본씨연중적구체공능급심입탐토DAD1단백재식물세포정서성사망중가능적작용궤제전정료기출。
[Objective] The aim was to clone NbDAD1 gene from Nicotiana benthami- ana and study its genetic transformation. [Method] NbDAD1 gene was isolated from N. benthamiana by using RT-PCR technology and over-expression vectors were con- structed to obtain NbDADl-overexpression resistant plants and NbDADl-overexpres- sion resistant plants carrying HA tag. [Result] The 351 bp long NbDAD1 gene was cloned from N. benthamiana; recombinant plasmids pCAMBIA1301-NbDAD1 and pCAMBIA1301-NbDAD1HAtag were constructed successfully; 50T0-generation N. ben- thamiana Hyg-resistant transgenic lines of three genotypes were obtained, including 23 positive transgenic plants. [Conclusion] This study laid the foundation for investi- gating the specific functions of NbDAD1 gene in N. benthamiana and exploring the possible functional mechanism of DAD1 protein in programmed cell death of plants.