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野油菜黄单胞菌葡萄糖激酶基因的克隆与表达 被引量：2

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摘要 葡萄糖激酶催化葡萄糖磷酸化生成葡萄糖-6-磷酸,是糖酵解的第一步反应,在野油菜黄单胞菌(Xcc)利用碳源代谢及其致病性中起重要作用。为了初步鉴定Xcc葡萄糖激酶基因功能,为研究其利用碳源相关代谢及致病性提供理论依据,以pQE30为表达载体,采用常规PCR技术对野油菜黄单胞菌Xcc8004基因组上注释为葡萄糖激酶的3个基因(XC_1166、XC_1223和XC_1976)进行了克隆与表达,该3个基因在大肠杆菌JM109中有较高表达量,经Ni2+金属层析柱纯化后分别得到带有3个His的融合蛋白XC_1166-His6、XC_1223-His6和XC_1976-His6,分析表明仅XC_1976的表达产物具有葡萄糖激酶活性,为Xcc8004葡萄糖激酶基因,经测定XC_1976-His6的最适反应温度为45℃,最适pH值为7.1,最适反应条件下比活力为(380.6±1.123)U/mg。
作者 杨正久 钱静 张连英 董红梅 陆光涛
机构地区 遵义医药高等专科学校医学技术系 广西大学生命科学与技术学院
出处 《江苏农业科学》 北大核心 2013年第6期14-17,共4页 Jiangsu Agricultural Sciences
基金 国家自然科学基金重点项目(编号:30130010) 国家自然科学基金面上项目(编号:30470944)
关键词 野油菜黄单胞菌 葡萄糖激酶 克隆 表达
分类号 Q75 [生物学—分子生物学]
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参考文献18

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二级参考文献27

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江苏农业科学
2013年 第6期

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