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OALib Journal期刊
ISSN: 2333-9721
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荧光假单胞菌高效广谱载体的构建及分析
DOI: 10.3724/SP.J.1145.2012.00292, PP. 292-297
Keywords: 荧光假单胞菌,slic,gc含量,表达载体,电转化,质粒稳定性
Abstract:
为促进高gc含量基因在荧光假单胞菌（pseudomonasfluorescens）中表达效果更加理想、操作更加简便，本研究首先采用不依赖基因序列和连接反应的克隆（sequenceandligationindependentcloning，slic）方法将载体pcibhis上与复制相关的序列和标记基因片段构建成克隆载体pcibs1.然后优化荧光假单胞菌转化方法，用电转化法将pcibs1导入荧光假单胞菌bl915中，随后又将t7和tac基因启动子分别插入pcibs1中，成功构建了表达载体pcibs3和pcibs2.研究发现载体pcibs1在大肠杆菌和荧光假单胞菌中均较为稳定，并且将绿色荧光蛋白基因插入表达载体中，在大肠杆菌bl21（de3）中获得表达，验证了表达载体功能.本研究构建的表达载体和建立的荧光假单胞菌bl915电转化方法，为高gc含量基因在荧光假单胞菌中的表达奠定了基础.图5表2参26
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