小麦蓝矮植原体SWP1效应蛋白的抗血清制备及检测应用  

Antiserum preparation and application of wheat blue dwarf phytoplasma effector SWP1

作　　者：羊海珍 王楠 吴薇 张子昂 吴云锋 YANG Hai-zhen;WANG Nan;WU Wei;ZHANG Zi-ang;WU Yun-feng(State Key Laboratory of Crop Stress Biology for Add Areas, Key Laboratory of Crop Pest Integrated Pest Management on Crop in Northwestern Loess Plateau, Ministry of Agriculture ,College of Plant Protection, Northwest A＆F University, Yangling 712100, China)

机构地区：[1]西北农林科技大学旱区作物逆境生物学国家重点实验室,农业部西北黄土高原作物有害生物综合治理重点实验室,西北农林科技大学植物保护学院,杨凌712100

出　　处：《植物病理学报》2018年第3期373-377,共5页Acta Phytopathologica Sinica

基　　金：国家自然科学基金(31570144;31371913)

摘　　要：小麦蓝矮病是西北冬麦区一种重要的植原体病害,造成严重的产量损失。为进一步探究小麦蓝矮植原体的致病机理,通过原核表达获得其致病因子SWP1的重组蛋白并制备抗血清。将PCR扩增到的SWP1基因片段插入到原核表达载体pET-30a(+)上,导入E.coli BL21(DE3)中,表达出约12 kDa含His标签的融合蛋白。用纯化的融合蛋白注射大白兔皮层,获得了特异性较强的SWP1抗血清,其效价为1∶4 000,并成功应用于SWP1在本氏烟中的检测。制备的抗血清在SWP1蛋白结构、功能及其与寄主的互作研究中具有重要意义。Wheat blue dwarf（ WBD） disease,a major phytoplasma disease of winter wheat,caused severe yield loss in northwestern China. To investigate the pathogenesis of WBD phytoplasma,WBD secreted protein SWP1 was expressed,purified,and used to produce an antibody. A 12 kDa SWP1 fusion protein was efficiently expressed in E. coli BL21（ DE3） with pET-30 a（ ＋） vector. The anti-SWP1 antibody was successfully applied to detect the transient expression of SWP1 in Nicotiana benthamiana. This work provides support for the further study on the structure of SWP1 and the interaction between SWP1 and its targets.

关 键 词：小麦蓝矮病植原体 SWP1 原核表达 抗血清 

分 类 号：S435.121[农业科学—农业昆虫与害虫防治]