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OALib Journal期刊
ISSN: 2333-9721
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苹果火疫解淀粉欧文氏usaw004菌株harpineausaw004基因的克隆和高效表达
DOI: 10.3724/SP.J.1145.2013.00986, PP. 986-989
Keywords: 欧文氏菌,克隆,harpineausaw004基因,harpineausaw004蛋白,植物过敏反应
Abstract:
从感染火疫病的苹果幼芽中分离纯化解淀粉欧文氏erwiniaamylovora菌株usaw004.该菌株能引起非寄主植物烟草叶片过敏反应.通过构建usaw004基因组文库和菌落原位杂交方法，从usaw004基因组文库中成功筛选到harpineausaw004基因的阳性克隆，并将其亚克隆到pbluescript-iisk(+).测定阳性克隆片段核苷酸序列harpineausaw004基因的orf长度为1212bp，转录产物harpineausaw004由403个氨基酸残基组成.将harpineausaw004基因克隆到表达载体pet28a(+)的t7启动子下，成功构建了高效表达质粒pt7-harpineausaw004，在大肠杆菌jm109中，经异丙基硫代-β-d半乳糖苷（iptg）诱导harpineausaw004基因高效表达，其表达产物可以诱导烟草叶片产生典型过敏反应症状.
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