绿竹锌指蛋白基因BoBZF克隆及功能初步分析

摘要

Zinc finger proteins play important roles in regulating gene expression in both prokaryote and eukaryotice. A zinc finger protein gene was isolated from leaves of Bambusa oldhamii with RT-PCR and RACE methods, and designed as BoBZF ( GenBank No. EU606025 ) . The full length cDNA of BoBZF is 1 076 bp containing an open reading frame which encodes 256 amino acids. The protein encoded by BoBZF has two B-Box domains, which indicates BoBZF belongs to B-Box zinc finger protein. Tissue specific expression showed that BoBZF expressed in leaf, sheath, stem and root, with the highest level in leaf. BoBZF gene was subcloned into the multiple cloning sites of pBI121 vector driven by 35S promoter, and transferred into Arabidopsis thaliana. RT-PCR analysis showed that BoBZF was expressed in the Arabidopsis. The transgenic plant was more drought tolerance than control, indicating that BoBZF was related to the drought tolerance ability of the bamboo.%锌指蛋白在原核生物与真核生物基因转录调控中发挥着重要作用.采用RT-PCR和RACE技术,从绿竹叶片中分离到1个锌指蛋白基因,命名为BoBZF(GenBank登记号:EU606025).BoBZF cDNA全长1 076 bp,编码256个氨基酸.蛋白结构分析表明,其编码的蛋白具有2个B-Box结构域,属于B-Box型锌指蛋白.组织特异性表达显示BoBZF在叶片、叶鞘、幼茎和根中均有表达,其中在叶片的表达丰度较高.将BoBZF置于CaMV 35S启动子控制下,构建到载体pBI121的多克隆位点,并转化拟南芥.RT-PCR分析表明,BoBZF已在拟南芥中表达.转基因植株耐旱性明显提高,意味着BoBZF与竹子的耐旱能力有关.