绿木霉Sm1基因的克隆、原核表达及蛋白纯化  

Sm1 gene cloning from Trichoderma virens,prokaryotic expression and protein purification

作　　者：朱洁伟[1] 吴琼[1] 陈捷[1] 

机构地区：[1]上海交通大学农业与生物学院、农业部都市农业(南方)重点开放实验室,上海200240

出　　处：《植物病理学报》2012年第3期334-336,共3页Acta Phytopathologica Sinica

基　　金：国家自然科学基金资助项目(30971949);教育部高校博士点专项基金(20090073110048);公益性行业(农业)科研专项(200903056);上海市科委重点项目(09391910900);上海市科委重大科技攻关项目(09dz1900103);现代农业产业技术体系专项(CARS-02)

摘　　要：木霉菌（Trichoderma spp.）是土壤内普遍存在的习居菌,具有多种生物防治功能,其中诱导植物免疫反应是重要的生物防治功能之一[1]。木霉菌Sm1蛋白（small one protein）是从绿木霉（Trichoderma virens）中分离得到的一种低分子量、富含半胱氨酸的疏水性蛋白,属于蛋白类激发子,与Cerato-platanin基因家族有很高的同源性,具有诱导植物免疫抗性的作用[2]。Trichoderma spp. have been recognized as the agents for the biocontrol of plant disease. They produce or release a variety of compounds that induce localized or systemic resistance responses in plants. In this study, Sml gene was cloned from Trichoderma virens （YZ2612）. Sequence analysis showed that the cDNA fragment had an ORF with length of 414 bp encoding 138 amino acid residues. The recombinant prokaryotic expression vector pET-28a（ ＋ ）-Sml was constructed, and then transformed into Escherichia coli BL21 （DE3）. IPTG concentration, induction time and temperature were optimized for enhancing fused protein expression, It was revealed that the best expression was achieved under conditions of 24°C and 1 mmol/L of IPTG for 4 hours. The recombinant protein was solubilized, denatured, refolded, and purified. These results might offer an important gene resource for being used to construct transgenic resistant lines of plants and to prepare new kind of protein biofungicide.

关 键 词：蛋白纯化 基因家族 绿木霉 原核表达 生物防治功能 克隆 植物免疫 小分子蛋白 

分 类 号：S511[农业科学—作物学]