小麦TaMAPK2激酶基因的原核表达及多克隆抗体-植物遗传资源学报.PDF

2011, 12( 1): 9295, 102 JournalofP lantGenetic Resources 小麦 TaMAPK2 激酶基因 的原核表达及多克隆抗体制备 1, 2 2 1 2 2 2 刘 沛 , 徐兆师, 晏月明,李连城 , 陈 明, 马有志 1 2 ( , 100048; / /, 100081) : MAP 蛋白激酶是一类重要的植物胁迫信号 控因子为了研究小麦 MAP 基因的功能, 本试验克隆了小麦 MAP 蛋白激酶基因 TaMAPK2为了制备TaMAPK2 基因的多克隆抗体, TaMAPK2 的非保守区段的 DNA序列 antiMAPK2 被 构建到原核表达载体 pET28a- ( + )上,表达融合蛋白 H isantiMAP 2在终浓度为 1mmol/L IPTG 诱导 1h的条件下,融合蛋 TM 白 H isantMi AP 2表达量达到最大通过蛋白标记亲和层析柱(H isTrap HP)得到纯化的 H isantMi AP 2融合蛋白利用新 西兰大白兔制备了 TaMAPK2 基因的多克隆抗体, ELISA竞争抑制法检测抗体效价检测,效价为 1: 80000,能满足后续试验要求 的效价值, 为进一步分析 TaMAPK2 的蛋白定位表达等提供基础 : 小麦;MAP 蛋白激酶; 原核表达;蛋白纯化; 多克隆抗体 Prokaryotic Expression and PolyclonalAntibody Preparation of the WheatTaMAPK2 Gene 1, 2 2 1 2 2 2 LIU Pei , XU Zhaoshi, YAN Yuem ing, LI Liancheng, CHENM ing ,MA Youzhi 1 2 ( College of Lfi e and H ealth Sciences, Cap ital N ormal Universit , B eij ing 100048; N a tionalK e Facilit f or C rop Gene R esources and Genetic Imp rovem ent /K e Labora tor of Crop Genetics and B reeding of M inistr of A griculture/Institute of Crop Sciences, ChineseA cad em of A gricu lture Scineces, B eij ing 100081) Abstract: MAP s are mi portant in stress signal transduction process of plant. In order to investigate the func tion ofwheatMAP s, aMAP genenamedTaMAPK2, was isolated from wheat. To obtain thepolydonalantibody of TaMAPK2, the nonconservation fraction ofTaMAPK2 genewas constructed into prokaryotic expression ve