Institute of Chemical Biology and Fundamental Medicine SB RAS, Lavrentieva Ave, 8, Novosibirsk, Russia.
Novosibirsk State University, Pirogova str, 2, Novosibirsk, Russia.
Extremophiles. 2019 Sep;23(5):599-612. doi: 10.1007/s00792-019-01119-2. Epub 2019 Aug 2.
A novel thermophilic bacteriophage AP45 and its host strain Aeribacillus sp. CEMTC656 were isolated from the Valley of Geysers, Kamchatka Peninsula, Russia. Bacteriophage AP45 was identified as a member of the Siphoviridae family by electron microscopy. It showed high thermostability and had a slow cycle of reproduction. The AP45 genome had 51,606 base pairs (bp) and contained 71 open reading frames (ORFs), 40 of them encoding proteins of predicted function. Genes encoding DNA and RNA polymerases were not identified, indicating that AP45 used host polymerases. Based on the ORF65 encoding putative endolysin, the recombinant protein rAP45Lys was developed and its peptidoglycan-hydrolyzing activity was demonstrated. The AP45 genome exhibited limited identity to other phage sequences; the highest identity, 36%, was with the genome of the thermophilic Geobacillus myovirus D6E. The majority of putative proteins encoded by the AP45 genome had higher similarity to proteins from bacteria belonging to the Bacillaceae family, than to bacteriophages. In addition, more than half of the putative ORFs in the AP45 genome were highly similar to prophage sequences of A. pallidus strain 8m3, which was isolated in north-east China. The AP45 phage and revealed prophages might be members of a new genus belonging to the Siphoviridae family.
一种新型嗜热噬菌体 AP45 及其宿主 Aeribacillus sp. CEMTC656 从俄罗斯堪察加半岛间歇泉谷中分离得到。电镜观察结果表明,噬菌体 AP45 属于长尾噬菌体科。它具有较高的热稳定性和缓慢的繁殖周期。AP45 基因组长 51606 个碱基对(bp),包含 71 个开放阅读框(ORFs),其中 40 个编码具有预测功能的蛋白质。未鉴定出编码 DNA 和 RNA 聚合酶的基因,表明 AP45 使用宿主聚合酶。根据编码潜在溶菌酶的 ORF65,开发了重组蛋白 rAP45Lys 并证明了其肽聚糖水解活性。AP45 基因组与其他噬菌体序列的同一性有限;与嗜热芽孢杆菌噬菌体 D6E 的基因组同源性最高,为 36%。AP45 基因组编码的大多数假定蛋白与芽孢杆菌科细菌的蛋白质具有更高的相似性,而与噬菌体的相似性较低。此外,AP45 基因组中超过一半的假定 ORF 与在中国东北地区分离的 A. pallidus 8m3 菌株的原噬菌体序列高度相似。AP45 噬菌体和揭示的原噬菌体可能属于长尾噬菌体科的一个新属。