Derzelle S, Hallet B, Francis K P, Ferain T, Delcour J, Hols P
Unité de Génétique, Université Catholique de Louvain, Louvain-La-Neuve, Belgium.
J Bacteriol. 2000 Sep;182(18):5105-13. doi: 10.1128/JB.182.18.5105-5113.2000.
An inverse PCR strategy based on degenerate primers has been used to identify new genes of the cold shock protein family in Lactobacillus plantarum. In addition to the two previously reported cspL and cspP genes, a third gene, cspC, has been cloned and characterized. All three genes encode small 66-amino-acid proteins with between 73 and 88% identity. Comparative Northern blot analyses showed that the level of cspL mRNA increases up to 17-fold after a temperature downshift, whereas the mRNA levels of cspC and cspP remain unchanged or increase only slightly (about two- to threefold). Cold induction of cspL mRNA is transient and delayed in time as a function of the severity of the temperature downshift. The cold shock behavior of the three csp mRNAs contrasts with that observed for four unrelated non-csp genes, which all showed a sharp decrease in mRNA level, followed in one case (bglH) by a progressive recovery of the transcript during prolonged cold exposure. Abundance of the three csp mRNAs was also found to vary during growth at optimal temperature (28 degrees C). cspC and cspP mRNA levels are maximal during the lag period, whereas the abundance of the cspL transcript is highest during late-exponential-phase growth. The differential expression of the three L. plantarum csp genes can be related to sequence and structural differences in their untranslated regions. It also supports the view that the gene products fulfill separate and specific functions, under both cold shock and non-cold shock conditions.
一种基于简并引物的反向PCR策略已被用于鉴定植物乳杆菌中冷休克蛋白家族的新基因。除了之前报道的两个cspL和cspP基因外,第三个基因cspC已被克隆并进行了表征。所有这三个基因都编码66个氨基酸的小蛋白,它们之间的同一性为73%至88%。比较Northern印迹分析表明,温度下降后,cspL mRNA的水平增加了17倍,而cspC和cspP的mRNA水平保持不变或仅略有增加(约两到三倍)。cspL mRNA的冷诱导是短暂的,并且根据温度下降的严重程度在时间上延迟。这三种csp mRNA的冷休克行为与四个不相关的非csp基因的情况形成对比,这四个基因的mRNA水平均急剧下降,在一种情况下(bglH),在长时间冷暴露期间转录本逐渐恢复。还发现这三种csp mRNA的丰度在最佳温度(28℃)下生长期间也有所不同。cspC和cspP的mRNA水平在延迟期最高,而cspL转录本的丰度在指数后期生长期间最高。植物乳杆菌的这三个csp基因的差异表达可能与其非翻译区的序列和结构差异有关。这也支持了这样一种观点,即在冷休克和非冷休克条件下,基因产物都具有各自独立的特定功能。