Franchini Alessandro G, Egli Thomas
Swiss Federal Institute for Aquatic Science and Technology, PO Box 611, Überlandstrasse 133, CH-8600 Dübendorf, Switzerland.
Microbiology (Reading). 2006 Jul;152(Pt 7):2111-2127. doi: 10.1099/mic.0.28939-0.
Microarray technology was used to study the cellular events that take place at the transcription level during short-term (physiological) and long-term (genetic) adaptation of the faecal indicator bacterium Escherichia coli K-12 to slow growth under limited nutrient supply. Short-term and long-term adaptation were assessed by comparing the mRNA levels isolated after 40 or 500 h of glucose-limited continuous culture at a dilution rate of 0.3 h(-1) with those from batch culture with glucose excess. A large number of genes encoding periplasmic binding proteins were upregulated, indicating that the cells are prepared for high-affinity uptake of all types of carbon sources during glucose-limited growth in continuous culture. All the genes belonging to the maltose (mal/lamB) and galactose (mgl/gal) operons were upregulated. A similar transcription pattern was observed for long-term cultures except that the expression factors were lower than in the short-term adaptation. The patterns of upregulation were confirmed by real-time RT-PCR. A switch from a fully operational citric acid cycle to the PEP-glyoxylate cycle was clearly observed in cells grown in glucose-limited continuous culture when compared to batch-grown cells and this was confirmed by transcriptome analysis. This transcriptome analysis confirms and extends the observations from previous proteome and catabolome studies in the authors' laboratory.
利用微阵列技术研究了粪便指示菌大肠杆菌K-12在有限营养供应下短期(生理)和长期(遗传)适应缓慢生长过程中转录水平上发生的细胞事件。通过比较在稀释率为0.3 h(-1)的葡萄糖限制连续培养40或500小时后分离的mRNA水平与葡萄糖过量的分批培养中的mRNA水平,评估短期和长期适应情况。大量编码周质结合蛋白的基因被上调,表明细胞在连续培养中葡萄糖限制生长期间为高亲和力摄取所有类型的碳源做好了准备。所有属于麦芽糖(mal/lamB)和半乳糖(mgl/gal)操纵子的基因均被上调。长期培养观察到类似的转录模式,只是表达因子低于短期适应。上调模式通过实时RT-PCR得到证实。与分批培养的细胞相比,在葡萄糖限制连续培养中生长的细胞中明显观察到从完全运行的柠檬酸循环向磷酸烯醇式丙酮酸-乙醛酸循环的转变,转录组分析证实了这一点。该转录组分析证实并扩展了作者实验室先前蛋白质组和分解代谢组研究的观察结果。