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丁香假单胞大豆致病变种harpin编码基因的克隆表达与功能研究

DOI: 10.7685/j.issn.1000-2030.2013.01.002, PP. 6-12

Keywords: 丁香假单胞大豆致病变种,harpin编码基因,GST融合表达,烟草,过敏性反应,促生

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Abstract:

采用PCR方法从丁香假单胞大豆致病变种(Pseudomonassyringaepv.glycinea)A1和S1菌株中分别克隆到大小为1026和1038bp的hrpZ基因(hrpZPsgA11和hrpZPsgS11),对该基因进行了原核表达和功能研究。SDS-PAGE显示其表达产物为相对分子质量62×103的融合蛋白,harpinZPsgA1和harpinZPsgS1的相对分子质量约为35×103。harpinZPsgS1的粗提蛋白经GSTrapFF纯化后质量浓度可达1.1mg?mL-1。生物活性检测表明,该蛋白对热稳定,对蛋白酶K敏感,可以在非寄主植物烟草上激发过敏反应,过敏反应可以被真核生物代谢抑制剂抑制,并且对烟草有明显的促生作用。序列比对发现,来自丁香假单胞大豆致病变种的hrpZ基因可分为2类,一类以hrpZPsgA1为代表,包括hrpZPsg12,另一类以hrpZPsgS1为代表,包括来自r0和Race4菌株的hrpZ基因。这2类hrpZ基因的核苷酸同源性为79%,氨基酸同源性为77%,均富含甘氨酸,不含半胱氨酸,与假单胞菌属以外的其他革兰氏阴性植物病原细菌harpin编码基因不存在相似性。

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