Gautam Aarti, Hathaway Marianne, Ramamoorthy Ramesh
Tulane National Primate Research Center, Division of Bacteriology and Parasitology, Tulane University Health Sciences Center, Covington, LA 70433, USA.
FEMS Microbiol Lett. 2009 Apr;293(2):278-84. doi: 10.1111/j.1574-6968.2009.01542.x. Epub 2009 Feb 26.
In this study, we investigated the functional elements of the flaB promoter of Borrelia burgdorferi. Promoter function was examined in a high-passage variant of strain JD1 using a set of 5' deletions and mutations within the flaB promoter. Expression from the modified flaB promoters was assayed using the gene for green fluorescent protein (gfp) as a reporter. Although the -35 element of the promoter stimulated promoter activity, its disruption did not negate expression. Sequences upstream of the -35 had no effect on expression. The -35/-10 spacer region composed of a T-rich sequence was critical for optimal promoter function. Surprisingly, a cytosine at the -13 site was found to be more favorable for transcription compared with a guanosine at the same site. Based on these results and other characteristics, we propose that the B. burgdorferi flaB promoter is an example of an extended -10 promoter. Further, the T-rich spacer is a key element of the flaB promoter that contributes to the abundance of the flagellar core protein in Borrelia species.
在本研究中,我们调查了伯氏疏螺旋体flaB启动子的功能元件。使用flaB启动子内的一组5'缺失和突变,在菌株JD1的高传代变体中检测启动子功能。使用绿色荧光蛋白(gfp)基因作为报告基因,检测修饰后的flaB启动子的表达。尽管启动子的-35元件刺激了启动子活性,但其破坏并未消除表达。-35上游的序列对表达没有影响。由富含T的序列组成的-35/-10间隔区对最佳启动子功能至关重要。令人惊讶的是,与同一位置的鸟苷相比,-13位点的胞嘧啶对转录更有利。基于这些结果和其他特征,我们提出伯氏疏螺旋体flaB启动子是延伸-10启动子的一个例子。此外,富含T的间隔区是flaB启动子的关键元件,有助于伯氏疏螺旋体物种中鞭毛核心蛋白的丰度。