萝卜过氧化物酶RNA干扰载体的构建及鉴定

Construct for Radish Peroxidase RNA Interference Vector and Identification

萝卜过氧化物酶在植物的生长、发育和防御等多种生理生化过程中发挥着重要作用。利用RNA干扰技术(RNA interference RNAi)来研究酶功能已日渐成熟。本文根据植物hpRNA(hairpin RNA)的原理,以萝卜过氧化物酶基因rsprx1序列为靶目标,选择rsprx1的保守区作为构建RNAi序列,借助中间克隆载体,经过3次亚克隆,最后形成含rsprx1-RNAi表达盒的双元表达载体pART27-Y2,并转入农杆菌EHA105。

Peroxidases of radish play an important role in plant growth,development,defense and other physiological and biochemical processes.The RNA interference technology comes to maturity to study the function of enzymes.In this paper,in accordance with the principles of plant hpRNA,the radish peroxidase rsprx1 was as target gene,the conservative region of sequence rsprx1 as RNAi construct sequence.With intermediate cloning vector,dual expression vector pART27-Y2 with rsprx1-RNAi expression cassette was made by subcloned three times and transfer it to Agrobacterium tumefaciens EHA105.