Zhao Miao, Gilbert Kerrigan, Danelishvili Lia, Jeffrey Brendan, Bermudez Luiz E
Program of Biology, Oregon State University, Corvallis, OR, USA.
Department of Biomedical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, OR, USA.
Adv Microbiol. 2016 Nov;6(13):927-941. doi: 10.4236/aim.2016.613087. Epub 2016 Nov 9.
is an opportunistic bacterium associated with pathogenic behavior in both humans and animals. has evolved as a pathogen by having an environmental component in its life style. Prophages are the integrated viral forms in bacterium genome. They constitute about 10% - 20% of genome of many bacteria and they contribute to pathogenicity of microbes. We investigated whether the 104 genome contained prophages and evaluated the genes/proteins for putative functions. Three prophage genes were identified in the 104 database, and sequences were analyzed for specific motifs. The prophage sequences were then cloned in and the bacterial phenotype was evaluated in gain of function assays for environmental stresses, such as tolerance to extreme temperatures, UV light, biofilm formation and resistance to acid as well as macrophage survival. The results indicate that two of the prophage genes, MAV_0696 and MAV_2265, confer with enhanced ability to produce biofilm. Using a Real-Time PCR, it was determined that MAV_0696 and MAV_2265 transcripts were upregulated upon biofilm formation by . The expression of MAV_2265 gene was significantly higher at all selected time points. In addition, the expression of MAV_2265 in . also led to significantly greater survival rate at pH 5.0 compared to the wild-type control. None of the other physical abilities were altered by overexpressing the prophage genes in . In summary, we identified three prophage sequences in 104, from which two of them were found to be associated with biofilm formation and one with resistance to the acidic environment. Future studies will identify the mechanisms involved in the prophages function.
是一种与人类和动物致病行为相关的机会致病菌。它通过在生活方式中具有环境成分而进化成为病原体。前噬菌体是细菌基因组中的整合病毒形式。它们约占许多细菌基因组的10% - 20%，并有助于微生物的致病性。我们研究了104基因组是否包含前噬菌体，并评估了这些基因/蛋白质的假定功能。在104数据库中鉴定出三个前噬菌体基因，并对序列进行了特定基序分析。然后将前噬菌体序列克隆到[具体载体]中，并在功能获得试验中评估细菌表型，以检测其对环境压力的耐受性，如对极端温度、紫外线、生物膜形成、耐酸性以及在巨噬细胞中的存活能力。结果表明，两个前噬菌体基因MAV_0696和MAV_2265赋予了增强的生物膜形成能力。使用实时PCR测定，发现MAV_0696和MAV_2265转录本在[具体细菌名称]形成生物膜时上调。MAV_2265基因在所有选定时间点的表达均显著更高。此外，与野生型对照相比，MAV_2265在[具体细菌名称]中的表达还导致在pH 5.0时存活率显著提高。在[具体细菌名称]中过表达前噬菌体基因并未改变其他任何生理能力。总之，我们在104中鉴定出三个前噬菌体序列，其中两个与生物膜形成相关，一个与耐酸性环境相关。未来的研究将确定前噬菌体功能所涉及的机制。
