小麦TaTCTP蛋白的原核表达及抗体制备  

Prokaryotic expression and antibody preparation of wheat TaTCTP protein

作　　者：麻楠 孙天杰[1] 刘超 焦园园 王冬梅[1] MA Nan;SUN Tianjie;LIU Chao;JIAO Yuanyuan;WANG Dongmei(State Key Laboratory of North China Crop Improvement and Regulation/Key Laboratory of Hebei Province for Plant Physiology and Molecular Pathology/College of Life Sciences,Hebei Agricultural University,Baoding 071001,China)

机构地区：[1]华北作物改良与调控国家重点实验室/河北省植物生理与分子病理学重点实验室/河北农业大学生命科学学院,河北保定071001

出　　处：《河北农业大学学报》2020年第4期10-15,29,共7页Journal of Hebei Agricultural University

基　　金：国家自然科学基金项目(31171472;31871548);河北省应用基础研究计划重点基础研究项目(12967149D).

摘　　要：为了阐明翻译控制肿瘤蛋白(Translationally controlled tumor protein,TCTP)在小麦-叶锈菌互作中的作用及其机制,本研究克隆了TaTCTP的开放阅读框(ORF)全长,并经过原核表达、亲和纯化、动物免疫,制备了TaTCTP兔多克隆抗体。结果表明,该基因CDS区全长为507 bp,编码168个氨基酸,预期分子量为18.8 kD,并具有较高的可溶性。以纯化后的TaTCTP重组蛋白制备的抗体可以特异性识别TaTCTP,效价为1∶6400。Western blotting检测发现TaTCTP在叶锈菌侵染后受诱导上调。本研究为进一步研究TaTCTP在小麦与叶锈菌互作过程中的功能及其作用机制奠定了基础。In order to study the function and mechanism of translationally controlled tumor protein(TCTP)in the interaction of wheat and P.triticina,the full-length open reading frame(ORF)of TaTCTP was cloned.After purification and animal immunization,TaTCTP rabbit polyclonal antibody was prepared.The results showed that the CDS region of this gene was 507 bp,encoding 168 amino acids,and had an expected molecular weight of 18.8 kD with high solubility.The antibody prepared with the purified TaTCTP recombinant protein specifically recognized TaTCTP with a titer of 1:6400.Western blotting detection revealed that TaTCTP was induced by P.triticina infection.This study laid a foundation for further study on the function and mechanism of TaTCTP in the interaction between wheat and P.triticina.

关 键 词：小麦 TCTP 原核表达 多克隆抗体 Western blotting 

分 类 号：S512.1[农业科学—作物学] Q943.2[生物学—植物学]