Detection of the outer membrane lipoprotein I and its gene in fluorescent and non-fluorescent pseudomonads: implications for taxonomy and diagnosis.

D De Vos, A Lim, P De Vos, A Sarniguet, K Kersters, P Cornelis
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引用次数: 42

Abstract

The open reading frame of the OprI lipoprotein gene from Pseudomonas aeruginosa was amplified by polymerase chain reaction (PCR) starting from purified DNA or colony lysates. A fragment of the expected size (249 bp) was detected in all P. aeruginosa strains from various clinical and geographical origins. The gene could only be amplified in pseudomonads of rRNA group I which are considered to be the authentic genus Pseudomonas. Digestions with HaeIII, PvuII and SphI of the amplified fragments demonstrated a sequence variation in the oprI gene. Colony, dot and Western blots with two monoclonal antibodies (mAbs) against the lipoprotein I confirmed our PCR results. These findings open interesting perspectives for the molecular taxonomy of the genus Pseudomonas and the development of diagnostic tools.

荧光和非荧光假单胞菌外膜脂蛋白I及其基因的检测:对分类和诊断的意义。
采用聚合酶链反应(PCR)从纯化的DNA或菌落裂解物开始扩增铜绿假单胞菌OprI脂蛋白基因的开放阅读框。在不同临床和地理来源的所有铜绿假单胞菌菌株中检测到预期大小的片段(249bp)。该基因只能在rRNA I群的假单胞菌中扩增,被认为是真正的假单胞菌属。对扩增片段HaeIII、PvuII和SphI进行消化,发现oprI基因序列发生变异。用两种针对脂蛋白I的单克隆抗体(mab)进行菌落、斑点和Western blots检测,证实了我们的PCR结果。这些发现为假单胞菌属的分子分类学和诊断工具的发展开辟了有趣的前景。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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