University of Nottingham, UK.
Curr Microbiol. 2010 Jun;60(6):419-27. doi: 10.1007/s00284-009-9559-8. Epub 2009 Dec 20.
We have transcriptionally profiled the genes differentially expressed in E. coli prey cells when predatorily attacked by Bdellovibrio bacteriovorus just prior to prey cell killing. This is a brief, approximately 20-25 min period when the prey cell is still alive but contains a Bdellovibrio cell in its periplasm or attached to and penetrating its outer membrane. Total RNA was harvested and labelled 15 min after initiating a semi-synchronous infection with an excess of Bdellovibrio preying upon E. coli and hybridised to a macroarray spotted with all predicted ORFs of E. coli. SAM analysis and t-tests were performed on the resulting data and 126 E. coli genes were found to be significantly differentially regulated by the prey upon attack by Bdellovibrio. The results were confirmed by QRT-PCR. Amongst the prey genes upregulated were a variety of general stress response genes, potentially "selfish" genes within or near prophages and transposable elements, and genes responding to damage in the periplasm and osmotic stress. Essentially, the presence of the invading Bdellovibrio and the resulting damage to the prey cell elicited a small "transcriptional scream", but seemingly no specific defensive mechanism with which to counter the Bdellovibrio attack. This supports other studies which do not find Bdellovibrio resistance responses in prey, and bodes well for its use as a "living antibiotic".
我们对捕食性攻击大肠杆菌的蛭弧菌在杀死宿主细胞前的差异表达基因进行了转录谱分析。这是一个短暂的过程,大约持续 20-25 分钟,此时宿主细胞仍然存活,但在其周质中含有蛭弧菌细胞,或附着并穿透其外膜。在半同步感染开始后 15 分钟,我们收获总 RNA 并进行标记,并用过量的蛭弧菌捕食大肠杆菌,然后将其杂交到带有大肠杆菌所有预测 ORF 的宏阵列上。对所得数据进行 SAM 分析和 t 检验,发现 126 个大肠杆菌基因受到蛭弧菌攻击的显著差异调控。通过 QRT-PCR 对结果进行了验证。上调的宿主基因包括各种一般应激反应基因、潜在的位于或靠近噬菌体和转座元件内的“自私”基因,以及对周质损伤和渗透压应激作出反应的基因。本质上,入侵的蛭弧菌的存在以及由此对宿主细胞造成的损伤引发了一个小的“转录尖叫”,但似乎没有特定的防御机制来对抗蛭弧菌的攻击。这支持了其他研究的结果,即宿主中不存在蛭弧菌的抗性反应,并为其作为“活体抗生素”的应用提供了良好的前景。