水稻光敏色素B抗体的制备及特异性检测  

Preparation of Rice Phytochrome B Antibody and Its Specificity Analysis

作　　者：闫丽华[1,2] 卢楠楠[1,2] 和亚男 尹海波[1] 谢先芝 

机构地区：[1]烟台大学生命科学学院,山东烟台264005 [2]山东省水稻研究所,山东济南250100

出　　处：《山东农业科学》2017年第8期1-6,共6页Shandong Agricultural Sciences

基　　金：山东省农业科学院农业科技创新工程项目(CXGC2016C01)

摘　　要：光敏色素是植物重要的光受体之一,主要感受红光和远红光。水稻光敏色素基因家族包括3个成员,即PHYA、PHYB和PHYC。我们已有研究表明PHYB在水稻生长发育和胁迫反应中具有重要作用。然而目前缺乏有效的水稻PHYB蛋白质抗体,极大地制约着PHYB基因作用机制的解析。在本研究中,我们比较了水稻光敏色素家族3个成员氨基酸序列同源性,选取了PHYB蛋白质C端的特异多肽。在此基础上,分离了特异多肽对应的c DNA序列,将其克隆在原核表达载体p ET16b上,利用异丙基硫代-β-D-半乳糖昔(IPTG)诱导、纯化获得了PHYB蛋白质特异性抗原,通过免疫新西兰大白兔,制备了PHYB蛋白的多克隆抗体。免疫印迹检测表明该抗体具有很高的特异性。本研究为深入解析PHYB基因在水稻生长发育中的作用机制提供了必要保障。Phytochrome（ PHY） is one of the important photoreceptors in plant. Phytochrome family mainly senses red and far-red lights. Rice phytochrome gene family is composed of three members known as PHYA,PHYB,and PHYC. Our previous studies has suggested that PHYB plays important roles in regulating rice growth and development as well as stress responses. However,absence of effective PHYB antibody greatly hinders the dissection of mechanisms of PHYB functions in rice. In this study,we selected the peptide specific to PHYB protein by comparing the amino acid identity among three rice phytochrome members. The c DNA fragment encoding specific PHYB peptide was isolated and cloned into prokaryotic expression vector p ET16 b.The PHYB antigen was obtained after isopropyl thio-beta-d-galactose（ IPTG） induction and purification of expressed protein. The polyclonal PHYB antibody was prepared by immunizing New Zealand rabbits. Western blotting assays revealed that PHYB polycolonal antibody prepared in this study was specific to detect PHYB protein in rice extracts. The results from this study were necessary for comprehensively exploring the functional mechanism of PHYB in rice.

关 键 词：水稻 光敏色素 原核表达 抗体 

分 类 号：S511[农业科学—作物学] Q78[生物学—分子生物学]