Wang Xing-Guo, Lin Bo, Kidder J Michael, Telford Samuel, Hu Linden T
Division of Geographic Medicine and Infectious Diseases, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
J Bacteriol. 2002 Nov;184(22):6198-206. doi: 10.1128/JB.184.22.6198-6206.2002.
We analyzed expression of a putative oligopeptide permease (Opp) of Borrelia burgdorferi. Unlike the opp operons of other bacteria for which there is a single substrate binding protein, B. burgdorferi codes for three substrate binding proteins (OppA-I to -III) in its opp operon and an additional two homologs on plasmids (OppA-IV and -V). Instead of a single promoter region regulating transcription of the entire operon, as seen in other bacterial opp operons, it appears that among oppA-I, -II, and -III, as well as oppA-IV and -V, each has a potential upstream promoter region. We tested the function of these putative promoter sequences by fusion to a promoterless beta-galactosidase reporter gene in pCB182. Each of the promoter regions was found to be active. The level of activity in the reporter constructs closely paralleled the level of expression of each gene in in vitro-grown B. burgdorferi. Changes in carbon and nitrogen availability differentially affected individual promoters, but no changes in promoter activity were seen when Escherichia coli bacteria (with the promoter constructs) were grown in various concentrations of phosphate and leucine and changes in pH. Expression of specific oppA genes with B. burgdorferi varied significantly between its mouse and fed and unfed tick hosts. Differences in regulation of opp gene expression suggest a potential role in environmental response by the organism.
我们分析了伯氏疏螺旋体假定的寡肽通透酶(Opp)的表达情况。与其他细菌的opp操纵子不同,其他细菌的opp操纵子只有一种底物结合蛋白,而伯氏疏螺旋体在其opp操纵子中编码三种底物结合蛋白(OppA-I至-III),并且在质粒上还有另外两种同源物(OppA-IV和-V)。与其他细菌opp操纵子中由单个启动子区域调控整个操纵子转录不同,在OppA-I、-II和-III以及OppA-IV和-V中,似乎每个都有一个潜在的上游启动子区域。我们通过将这些假定的启动子序列与pCB182中无启动子的β-半乳糖苷酶报告基因融合,来测试其功能。发现每个启动子区域都是有活性的。报告构建体中的活性水平与体外培养的伯氏疏螺旋体中每个基因的表达水平密切平行。碳源和氮源可用性的变化对各个启动子有不同影响,但当含有启动子构建体的大肠杆菌在不同浓度的磷酸盐、亮氨酸以及pH值变化的条件下生长时,启动子活性未见变化。伯氏疏螺旋体中特定oppA基因的表达在其小鼠宿主以及饱血和未饱血蜱宿主之间存在显著差异。opp基因表达调控的差异表明该生物体在环境应答中可能发挥作用。