酵母双杂交系统筛选与AtTR1相互作用的蛋白

Screening of candidate proteins interacting withAtTR1 by yeast two-hybrid system

AtTR1是拟南芥中发现的与植物高温和高盐等非生物胁迫相关的全新基因.采用GAL4酵母双杂交系统,以AtTR1作为诱饵蛋白,对拟南芥cDNA文库进行筛选,获得了与AtTR1有潜在相互作用的蛋白质PATL6(Patellin6).进一步采用GST pull-down方法在体外验证了PATL6与AtTR1的相互作用.采用Real-time PCR检测PATL6在NaCl和Mannitol处理后的表达情况,结果显示PATL6的表达受高盐和高渗的诱导,是一个与非生物胁迫应答相关的基因.这些实验结果为进一步研究AtTR1及其相互作用蛋白质在植物抗逆胁迫中的作用奠定了基础.

AtTR1 was a new gene which is relative to high temperature and salt stress. AtTR1 was used as a bait to screen proteins interacting with AtTR1 in Arabidopsis cDNA library by yeast two-hybrid system. PATL6 （Patellin6） which could interact with AtTR1 was obtained. GST pull-down result showed that PATL6 could interact with AtTR1 in vitro. The expression of PATL6 in Arabidopsis after treated with NaC1 and Mannitol was assayed by Real-time PCR. The result showed that the expression level could be induced by high salt and high osmotic stresses and PATL6 was an abiotic stress response- relate gene. This study provides the foundation for further understanding the functions of these proteins in plant defense responses.