prd29A-otsAB表达载体的构建与转化本生烟草的研究

摘要

分别以拟南芥(Arabidopsis thaliana)和大肠杆菌(Escherichia coli)的基因组DNA为模板,通过PCR技术克隆得到rd29A启动子、otsA和otsB基因,将其依次插入到p2300-gfp质粒中,从而构建p2300-prd29A-otsAB融合基因表达载体.利用农杆菌介导法将prd29A-otsAB融合基因导入到本生烟草(Nico-tiana benthamiana)中,通过对转基因烟草进行筛选,初步获得具有卡那霉素抗性的转基因植株,为今后进一步研究相关基因的功能以及通过基因工程技术提高植物的抗胁迫能力奠定基础.%The rd29A promoter of Arabidopsis thaliana,otsA and otsB genes in Escherichia coli were cloned by PCR respec-tively. Afterwards,the target fragments were ligated into plasmid p2300-gfp,which would lead to the construction of p2300-prd29A-otsAB expression vector. By Agrobacterium-mediated method,the prd29A-otsAB fusion gene was introduced into Nico-tiana benthamiana,and then the transgenic Nicotiana benthamiana was obtained through the tissue culture and antibiotics screening for the first time. The experimental results can provide the reference for improving the stress resistance of plants through genetic engineering.