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OALib Journal期刊
ISSN: 2333-9721
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干燥和复吸水条件下发菜过氧化物氧还蛋白差异表达与基因克隆
DOI: 10.3724/SP.J.1145.2011.00666, PP. 662-672
Keywords: 发菜,过氧化物氧还蛋白,双向电泳,差异表达,基因克隆,原核表达,耐旱
Abstract:
发菜（nostocflagelliforme）是一种陆生固氮蓝藻，为探讨其耐旱的分子机理及对极端干旱环境的适应和保护机制，运用双向电泳技术（2-de）、凝胶图像分析、maldi-tof-tof/ms质谱鉴定和数据库检索，分析发菜过氧化物氧还蛋白（peroxiredoxin）在持续干燥48h和复吸水4h后的差异表达水平，根据鉴定的过氧化物氧还蛋白已知氨基酸序列设计简并性引物克隆该基因，分析基因和氨基酸序列的同源性及对蛋白质二级结构进行预测，并研究其原核表达.结果表明，发菜过氧化物氧还蛋白在复吸水后的表达量明显高于干燥状态下的表达量。根据简并性引物克隆获得长度为639bp的过氧化物氧还蛋白基因，genbank登陆号为hm854286.序列比较分析显示该基因具有较高的保守性，蛋白质二级结构主要由α螺旋、β折叠和随机卷曲构成.将过氧化物氧还蛋白基因在大肠杆菌中表达，获得符合预期的外源重组蛋白（26.5×103），经westernblotting验证，该外源蛋白为过氧化物氧还蛋白.图10表1参25
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