核盘菌多聚半乳糖醛酸酶基因的克隆及hpRNAi载体的构建

Molecular Cloning of Polygalacturonase Gene from Sclerotinia Sclerotiorum and the Construction of its Corresponding hpRNAi Vector

多聚半乳糖醛酸酶是核盘菌致病力发挥作用的关键酶。本研究用PCR的方法从核盘菌基因组DNA中扩增出多聚半乳糖醛酸酶(PG5)基因片段,再以扩增出的PG5基因片段作为模板设计引物扩增出一个相应较小的片段。将两个PG5基因片段反向插入到植物表达载体2300 sn的35S启动子和nos终止子之间,构建出可转录表达出发夹RNA结构的组成型油菜RNA干扰载体,为今后油菜利用RNA干扰抗菌核病的基因工程研究奠定了基础。

Polygalacturonase plays important roles in the way of Sclerotinia sclerotiorum＇s exerting ability of caused the death. From the point of view of genetic engineering, polygalacturonase （ PG5 ） was cloned by PCR technique from Sclerotinia sclerotiorum genomic DNA. The two fragments were ligated in opposite directions into plant expression vector 2300sn, and respectively to produce hpRNAi vector 2300PG. This is the first step to insist Sclerotinia sclerotiorum in rapeseed by using RNA interference.