2 Escehrichia hermannii Wzt遺伝子転写量のreal time quantitative RT-PCR法による解析(第513回大阪歯科学会例会)

概要

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We recently reported that wzt encoding an ATP binding protein in the O-antigen biosynthesis and export system was crucial for the biofilm formation of Escherichia hermannii. In this study, we attempted to establish the real time reverse transcription polymerase chain reaction (RT-PCR) conditions necessary to evaluate the transcriptional level of wzt along with growth of biofilm-positive and -negative strains of this organism. Transcriptional levels of wzt were normalized with a transcriptional level of the 16S rRNA gene. The relative expression levels of wzt were compared between the biofilm-positive wild type strain YS-11, the wzt-disrupted mutant 455 and the wzt-overexpressed recombinant strain 455-LM, We confirmed by melting curve analysis that the real time RT-PCR condition used in this study amplified the target gene. Also, we found that the relative expression level of wzt was greatest in strain 455-LM, while it was expressed to a lesser extent in YS-11, among the strains used. Our results suggest that this method can be used to evaluate relative transcriptional levels of wzt in E. hermannii.
2008-06-25