Ca2 是非生物胁迫信号转导途径中的重要信号分子,植物类钙调磷酸酶B亚基蛋白(calcineurin B-like proteins,CBL)是一类重要的钙信号受体蛋白,主要通过与其他蛋白的特异结合将信号传递下去,使植物形成对非生物胁迫的响应。本实验室已经获得大豆GmCBL1基因,功能鉴定显示GmCBL1增强了转基因拟南芥对非生物胁迫的耐性。为了进一步研究GmCBL1的作用机理,本研究构建诱饵载体pGBKT7-GmCBL1,利用酵母双杂交实验技术,将pGBKT7-GmCBL1及pGADT7和大豆cDNA 文库混匀转入AH109 酵母感受态细胞,筛选大豆GmCBL1的互作蛋白。通过对筛选获得的106个蛋白基因测序和Blast比对分析结果,并根据其可能的生理功能对这些候选蛋白归类,主要整理得到四类蛋白:能量代谢相关蛋白、修饰蛋白、防御蛋白、钙信号转导相关蛋白。筛选得到候选蛋白的功能预测初步表明大豆GmCBL1参与多条信号途径,为进一步探索大豆CBL介导的抗逆信号途径打下基础。
Calcium is a critical messenger in abiotic stress responses signal transduction pathways in plants. Calcineurin B-like proteins (CBLs) represent a family of plant calcium sensor protein that function in calcium signaling by interacting with their interacting protein to transduce signal, making plant response to abiotic stress. Our laboratory has identifies a putative soybean CBL gene that functions in abiotic stress tolerance tolerance in the transgenic Arabidopsis plants. In order to further explore the GmCBL1-mediated stress resistance mechanism, the mixture of bait vectors pGBKT7-GmCBL1 were constructed, pGADT7 and soybean cDNA library was introduced into yeast competent cells AH109 to screen the interaction proteins from soybean cDNA library by yeast-two hybrid system. A total of 106 candidate positive clones were sequenced, analyzed through BLAST. According to their possible physiological function , those candidate proteins were classified into one of four categories: energy metabolism proteins, modified protein, defense protein, calcium signaling transduction proteins. Function prediction of the candidate proteins suggested that GmCBL1 was possibly involved in several stress signal transduction pathways.This work provides a basis for a defined further functional dissection of soybean CBL mediated signaling system.
李林蔚,张双喜,周永斌,等.大豆类钙调磷酸酶B亚基GmCBL1互作候选蛋白的筛选[J].植物遗传资源学报,2015,16(2):359-363.
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