稻瘟病菌MGG_14095基因克隆、表达及其表达产物的纯化

Cloning, Expression of MGG_14095 Gene from Magnaporthe oryzae and Purification of Its Translated Product

稻瘟病菌(Magnaporthe oryzae)是世界上危害性最大的水稻病原菌,也是阐明植物真菌病分子基础的主要模式生物,开展稻瘟病菌生长发育的相关研究,对稻瘟病的防治有重要意义。MGG_14095基因是稻瘟病菌的致病基因,对MGG_14095蛋白进行生物信息学预测,结果显示,MGG_14095蛋白等电点(pI)为5.72,不稳定指数为50.99,属于酸性不稳定蛋白;含角质酶保守结构域,隶属α/β水解酶超家族;有明显跨膜结构和信号肽剪切位点,属于分泌型蛋白质。本研究克隆稻瘟病菌MGG_14095(38-281)基因,构建原核表达系统pETM20-MGG_14095(38-281),采用分步层析纯化MGG_14095(38-281)蛋白,成功获得高纯度可溶目标蛋白。本研究结果为解析MGG_14095(38-281)蛋白质结构、探索稻瘟病菌致病机理及对稻瘟病菌防治提供一定的理论与试验依据。

Magnaporthe oryzae is the most harmful pathogen in the world for rice and the main model organism for elucidating the molecular basis of plant fungal disease. Carrying out relevant research of the growth and development of Magnaporthe oryzae has a great significance for the control of Magnaporthe oryzae. MGG_14095 gene is the pathogenic gene of Magnaporthe oryzae. The bioinformatics prediction of MGG_14095 protein showed that the isoelectric point(pI) of MGG_14095 protein was 5.72, and the instability index was 50.99, belonging to the acid unstable protein and the α/β hydrolase superfamily, containing the keratinase conserved domain;having obvious transmembrane structure and signal peptide cutting site, belonging to the secretory protein. In this study, we cloned the MGG-14095(38-281) gene of Magnaporthe oryzae, constructed the prokaryotic expression system pETM20-MGG_14095(38-281), used the stepwise chromatography to purify MGG_14095(38-281) protein, and the high-purity soluble target protein was obtained successfully. The results provided certain theoretical and experimental basis for the analysis of MGG_14095( 38-281) protein structure, the exploration and the control of pathogenetic mechanism of Magnaporthe oryzae.