摘要
本研究依据HVA1基因序列克隆六棱大麦HVA1基因cDNA片段,构建Ubiquitin启动子驱动下的植物表达载体pCAMBIA1300-HVA1。然后通过三亲杂交法将重组质粒PCAMBIA1300-HVA1转入农杆菌LBA4404,并采用农杆菌介导法转化烟草。经PCR,PCR-Southern blotting和RT-PCR检测表明HVA1基因已整合进烟草基因组,并在转录水平上获得表达。功能验证的结果显示,转基因植株叶片的保水率提高了近1倍,暗示转基因烟草具有一定的抗旱潜力。
In this article,the barley HVAlcDNA gene was cloned according to HVA1 sequence..And a botanic expression vector PCAMBIA1300-HVA1 which was driven by Ubiqnifin promoter was constructed. Then the recombinant plasmid was introduced into Agrobacterium tumefaciens LBA4404 by tri-parental mating and was transformed into tobacco with Agrobacterium tumefaciens-mediated. It was proved that HVAlcDNA gene had been integrated into the tobacco genome by PCR and PCR-Southren blotting analysis. It was confirmed that HVAlcDNA gene was expressed by RT-PCR at transcription level. The results of function validation indicated that the leaf water retaining capacity of transgenic tobacco plants was higher one fold than the control .It implied that there was drought-resistance potential for the transgenic tobacco plants.
出处
《生物技术通报》
CAS
CSCD
2007年第3期139-144,共6页
Biotechnology Bulletin
基金
农业部"948"项目资助(2003-3Q)
关键词
HVA1
根癌农杆菌
遗传转化
烟草
抗旱
HVA1 Agrobacterium tumefaciens Genetic transformation Tobacco Drought-resistance
作者简介
李楠（1982-），女，汉族，北京，在读硕士生，研究方向：植物基因工程
通讯作者：赵琦，女，教授，博士生导师，主要从事植物生理生化、分子生物学研究
