抗菌肽Rs-AFP_2的原核表达及抑菌活性  

Expression of Rs-AFP_2 in Escherichia coli and Its Antifungal Assay

作　　者：路妍[1,2] 刘宝业[2] 井金学[1] 张增艳[2] 

机构地区：[1]西北农林科技大学植物保护学院,陕西杨凌712100 [2]中国农业科学院作物科学研究所/农作物基因资源与基因改良国家重大科学工程,北京100081

出　　处：《植物遗传资源学报》2010年第1期84-88,共5页Journal of Plant Genetic Resources

基　　金：国家转基因生物新品种培育科技重大专项(2008ZX08002-001)

摘　　要：为了明确人工合成的萝卜抗菌肽Rs-AFP2基因编码产物是否对一些重要农作物病原真菌有抑制作用,本文将该基因编码序列亚克隆到原核表达载体pGEX-4T-1上,构建成GST-Rs-AFP2融合蛋白表达载体。经异丙基-β-D-硫代半乳糖苷(IPTG)诱导,获得了以包涵体形式表达的GST-Rs-AFP2重组蛋白。经过裂解、洗涤、溶解、复性、离心等处理,获得了纯化的GST-Rs-AFP2融合蛋白。对重要作物病原真菌进行体外抑菌分析的结果表明,Rs-AFP2可以显著抑制水稻纹枯病菌、小麦纹枯病菌、小麦根腐病菌、棉花黄萎病菌、小麦赤霉病菌的菌丝生长,说明Rs-AFP2可作为上述植物病害抗性基因育种的潜在基因。Rs-AFP2 is an antifungal peptide from radish （Raphanus sativus）. In this study, the full cDNA sequence of Rs-AFP2 gene was synthesized, and was sub-cloned in-frame to the 3 ＇-terminus of the ORF of Glutathione S-transferase（GST） in the prokaryotic expression vector pGEX-4T-1. This allowed the recombinant protein GST- Rs-AFP2to be highly expressed in cells of Escherichia coll. The recombinant protein GST-Rs-AFP2 could be expressed in the form of inclusion bodies after induction by isopropyl-13-D-thiogalactoside. Then, the GST-Rs-AFP2 protein was solubilized, denatured, refolded, and purified. The results of antifungal assay in vitro showed that the Rs- AFP2 protein exerted obvious inhibition against the growth of phytopathogenic fungi, such as Rhizoctonia solani,R. cerealis,Bipolaris sorokiniana, Verticilliulll dahliae, and Fusarium graminearum. Therefore, Rs-AFP2 gene may be useful for improving the crop resistance against the above pathogenic fungi.

关 键 词：抗菌蛋白Rs-AFP2 复性 抑制真菌 植物病原真菌 

分 类 号：S332[农业科学—作物遗传育种]