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RNA Arbitrarily Primed PCR and Fourier Transform Infrared Spectroscopy Reveal Plasticity in the Acid Tolerance Response of Streptococcus macedonicus▿

机译:RNA任意引物PCR和傅立叶变换红外光谱显示马其顿链球菌耐酸反应的可塑性

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摘要

We have previously reported that an acid tolerance response (ATR) can be induced in Streptococcus macedonicus cells at mid-log phase after autoacidification, transient exposure to acidic pH, or acid habituation, as well as at stationary phase. Here, we compared the transcriptional profiles of these epigenetic phenotypes, by RNA arbitrarily primed PCR (RAP-PCR), and their whole-cell chemical compositions, by Fourier transform infrared spectroscopy (FT-IR). RAP-PCR fingerprints revealed significant differences among the phenotypes, indicating that gene expression during the ATR is influenced not only by the growth phase but also by the treatments employed to induce the response. The genes coding for the mannose-specific IID component, the 1,2-diacylglycerol 3-glucosyltransferase, the 3-oxoacyl-acyl carrier protein, the large subunit of carbamoyl-phosphate synthase, and a hypothetical protein were found to be induced at least under some of the acid-adapting conditions. Furthermore, principal component analysis of the second-derivative-transformed FT-IR spectra segregated S. macedonicus phenotypes individually in all spectral regions that are characteristic for major cellular constituents like the polysaccharides of the cell wall, fatty acids of the cell membrane, proteins, and other compounds that absorb in these regions. These findings provide evidence for major changes in cellular composition due to acid adaptation that were clearly different to some extent among the phenotypes. Overall, our data demonstrate the plasticity in the ATR of S. macedonicus, which reflects the inherent ability of the bacterium to adjust the response to the distinctiveness of the imposed stress condition, probably to maximize its adaptability.
机译:我们以前曾报道过,自酸化,瞬时暴露于酸性pH或酸性习性以及稳定期后,在对数中期,可以在马其链球菌细胞中诱导耐酸反应(ATR)。在这里,我们比较了这些表观遗传表型的转录谱,通过RNA任意引物PCR(RAP-PCR),以及它们的全细胞化学组成,通过傅立叶变换红外光谱(FT-IR)。 RAP-PCR指纹图谱揭示了表型之间的显着差异,表明ATR期间的基因表达不仅受到生长期的影响,还受到诱导反应的处理方式的影响。发现至少诱导了编码甘露糖特异性IID成分,1,2-二酰基甘油3-葡萄糖基转移酶,3-氧代酰基酰基载体蛋白,氨基甲酰基磷酸合酶的大亚基和假设蛋白的基因。在某些酸适应条件下。此外,对二阶导数变换的FT-IR光谱进行主成分分析,可以在所有光谱区域中分别分离马其链球菌表型,这些特征是主要细胞成分的特征,例如细胞壁的多糖,细胞膜的脂肪酸,蛋白质,以及在这些区域吸收的其他化合物。这些发现为由于酸适应引起的细胞组成的重大变化提供了证据,这些变化在表型之间明显不同。总的来说,我们的数据表明马其顿菌的ATR具有可塑性,这反映了细菌固有的能力,即它可以调节对所施加压力条件的独特性的响应,从而最大程度地提高其适应性。

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