【目的】克隆Phytophthora capsici的elicitin基因并对其原核表达产物的生物活性进行研究。【方法】根据疫霉菌elicitin基因序列的保守性设计了2条引物,采用RT-PCR从P.capsici中克隆到长度为357bp的cDNA片段。【结果】将该cDNA在网上进行分析表明该基因的编码产物为P.capsici的elicitin,即capsicin;该蛋白质N端前20个氨基酸为信号肽,等电点为4.2。同时将该蛋白质的氨基酸序列与Genbank中登录的14个elicitin进行聚类分析,发现所克隆的elicitin与Phytophthora drechsleri的α-elicitin及Phytophthora megasperma的α-elicitin聚为一组,表明该elicitin蛋白质属于α-elicitin。经southern杂交分析表明,该基因在P.capsici基因组中最少存在两个拷贝。该基因的原核表达产物能诱导野生型烟草Xanthi产生过敏反应和对TMV与Phytophthora nicotianae产生系统获得抗性,且能诱导该烟草的PR-1a、PR-1b和PR-1c基因的表达;但是...
【Objective】We cloned a gene encoding elicitin from phytophthora capsici and elucidated the bioacitivity of prokaryotic expression of the gene. 【Method】A cDNA fragment of 357 bp encoding capsicin, was amplified from Phytophthora capsici by reverse transcript PCR (RT-PCR) based on high homology of elicitin genes from various Phytophthora spp. 【Result】The gene codes a predicted 118 amino acids protein containing a 20-amino acid signal peptide of that is cleaved before secretion and with a calculated pI of 4.2....