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Author:
Date:
2012-10-20
[Abstract] One major mechanism of phase variable gene expression in prokaryotes is through inversion of the promoter element for a gene or operon. This protocol describes how to detect the promoter orientation of a phase-variable gene by PCR. This protocol, including primer design, is specific to detection of the promoter orientations of hyxR, a LuxR-like response regulator in Extraintestinal Pathogenic Escherichia coli (ExPEC) isolates (Bateman and Seed, 2012); however, this protocol can be generalized to other organisms and genes to discriminate prokaryotic promoter inversions by PCR through size discrimination of the amplification products. Expression of hyxR is regulated through bidirectional phase inversion of the upstream promoter region mediated by a member of the family of ...
[摘要] 在原核生物中相变基因表达的一个主要机制是通过逆转基因或操纵子的启动子元件。该协议描述了如何通过PCR检测相变基因的启动子方向。该方案包括引物设计，其特异性用于检测hyxR（在肠道致病性大肠杆菌（ExPEC）分离株中的LuxR样应答调节剂）的启动子定向（Bateman和Seed，2012）;然而，该协议可以推广到其他生物体和基因以通过PCR通过扩增产物的大小鉴别来区分原核启动子逆转。 hyxR的表达通过由称为Fim样重组酶的位点特异性酪氨酸重组酶家族成员介导的上游启动子区的双向相位反转来调节。重组酶识别启动子侧翼的反向DNA重复序列并产生基因组重排，使启动子定向有利于或不利于基因表达。
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