摘要
根癌土壤农杆菌Ti质粒毒性区VirD1蛋白表现拓扑异构酶活性,能解除T-DNA边界处25bp重复序列的超螺旋状态,使外源基因的插入成为可能。本实验以C58菌株为模板,设计了一对含有BamHI和XhoI酶切位点的引物,成功克隆了VirD1基因,并构建了原核表达载体pET30a-VirD1,将其在宿主菌BL21中获得了表达,为将来构建含有VirD1基因的植物表达载体并协助目的基因转化奠定基础。
Agribacterium tumefaciens＇ Ti Plasmid VirD1 gene has topoisomerase activity. It can relieve the superhelical DNA at the 25bp reptition sequence of T-DNA boundary. Thus it is possible for outside genes to insert. In this study, according to the agribacterium tumefaciens＇ Ti plasmid gene sequences published by U. Washington we designed a pair of primers, which include Barn HI and Xho I, and clone the VirD1 gene correctly, constructed prokaryotic expression vectorpET30a-VirD1, and expressed in the host germ BL21 successfully. This study provides an available method and a theorical significance.
出处
《石河子大学学报（自然科学版）》
CAS
2005年第4期473-476,共4页
Journal of Shihezi University(Natural Science)
基金
兵团重点攻关项目(NKB.99SDXR02NY)
