Tóth István, Sváb Domonkos, Bálint Balázs, Brown-Jaque Maryury, Maróti Gergely
Institute for Veterinary Medical Research, Centre for Agricultural Research, Hungarian Academy of Sciences, Budapest, Hungary.
Seqomics Biotechnology Ltd., Mórahalom, Hungary.
Infect Genet Evol. 2016 Jan;37:150-7. doi: 10.1016/j.meegid.2015.11.022. Epub 2015 Nov 23.
Here we report the first complete nucleotide sequence of a Shiga toxin (Stx) converting phage from a Shigella sonnei clinical isolate that harbors stx1 operon, first identified in the chromosome of Shigella dysenteriae type 1. The phage named Shigella phage 75/02 Stx displayed Podoviridae morphology. It proved to be transferable to Escherichia coli K-12 strains, and cytotoxicity of the lysogenized strains was demonstrated in Vero cell cultures. Genomic analysis revealed that the prophage genome is circular and its size is 60,875 nt that corresponds to 76 ORFs. The genome of Shigella phage 75/02 Stx shows a great degree of mosaic structure and its architecture is related to lambdoid phages. All the deduced proteins, including the 37 hypothetical proteins showed significant homologies to Stx phage proteins present in databases. The phage uniformly inserted into the ynfG oxidoreductase gene framed by phage integrase and antirepressor genes in parental S. sonnei and in the three lysogenized K-12 strains C600, DH5α and MG1655. The Stx1 prophage proved to be stable in its bacterial hosts and remained inducible.
在此,我们报告了从一株携带stx1操纵子的宋内志贺氏菌临床分离株中获得的产志贺毒素(Stx)噬菌体的首个完整核苷酸序列,该操纵子最初在1型痢疾志贺氏菌染色体中被鉴定出来。名为志贺氏菌噬菌体75/02 Stx的噬菌体呈现出短尾噬菌体科的形态。它被证明可转移至大肠杆菌K-12菌株,并且在Vero细胞培养物中证实了溶原化菌株的细胞毒性。基因组分析显示,前噬菌体基因组是环状的,其大小为60,875 nt,对应76个开放阅读框。志贺氏菌噬菌体75/02 Stx的基因组呈现出高度的镶嵌结构,其结构与λ样噬菌体相关。所有推导的蛋白质,包括37个假定蛋白质,与数据库中存在的Stx噬菌体蛋白质显示出显著的同源性。该噬菌体在亲本宋内志贺氏菌以及三个溶原化的K-12菌株C600、DH5α和MG1655中均一致地插入到由噬菌体整合酶和抗阻遏基因框定的ynfG氧化还原酶基因中。Stx1前噬菌体在其细菌宿主中被证明是稳定的,并且仍然可诱导。