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幼虫芽孢杆菌新型噬菌体vB_PlaP_SV21的分离、特性鉴定及比较基因组分析

Isolation, characterization, and comparative genomic analysis of vB_PlaP_SV21, new bacteriophage of Paenibacillus larvae.

作者信息

Bozdeveci Arif, Akpınar Rahşan, Karaoğlu Şengül Alpay

机构信息

Biology Department, Faculty of Arts and Sciences, Recep Tayyip Erdogan University, Rize, Turkey.

Veterınary Control Instıtute, Bee Diseases, Samsun, Turkey.

出版信息

Virus Res. 2021 Nov;305:198571. doi: 10.1016/j.virusres.2021.198571. Epub 2021 Sep 27.

Abstract

Paenibacillus larvae cause an American foulbrood disease (AFB) that is responsible for the extinction of honeybee colonies and is a honeybee bacterial disease that has to be obligatory notified worldwide. Recently, bacteriophage studies targeting Paenibacillus larvae have emerged as a promising alternative treatment method. The inability of bacteria to create resistance against bacteriophages makes this method advantageous. As a consequence, this study was conducted to describe the genome and biological characteristics of a novel phage capable of lysing Paenibacillus larvae samples isolated from honeybee larva samples in Turkey. The Paenibacillus phage SV21 (vB_PlaP_SV21) was isolated by inducing Paenibacillus larvae strain SV21 with Mitomycin-C. Whole-genome sequencing, comparative genomics, and phylogenetic analysis of vB_PlaP_SV21 were performed. Transmission electron microscopy images showed that vB_PlaP_SV21 phage was a Podovirus morphology. The vB_PlaP_SV21 phage specific for Paenibacillus larvae was determined to belong to the Podoviridae family. Host range and specificity, burst size, lytic activity, and morphological characteristics of the phage were determined. Bioinformatic analysis of the Paenibacillus phage SV21 showed 77 coding sequences in its linear 44,949 bp dsDNA genome with a GC content of 39.33%. In this study, we analysed the genomes of all of the currently sequenced P. larvae phage genomes and classified them into five clusters and a singleton. According to molecular, morphological, and bioinformatics results, ıt was observed that API480 (podovirus), which was reported as a singleton in previous studies and public databases, and Paenibacillus phage SV21 phage could form a new cluster together.

摘要

幼虫芽孢杆菌会引发美洲幼虫腐臭病(AFB),这种疾病会导致蜂群灭绝,是一种必须在全球范围内强制通报的蜜蜂细菌性疾病。最近,针对幼虫芽孢杆菌的噬菌体研究已成为一种有前景的替代治疗方法。细菌无法对噬菌体产生抗性,这使得该方法具有优势。因此,本研究旨在描述一种新型噬菌体的基因组和生物学特性,该噬菌体能够裂解从土耳其蜜蜂幼虫样本中分离出的幼虫芽孢杆菌样本。通过用丝裂霉素-C诱导幼虫芽孢杆菌菌株SV21,分离出了幼虫芽孢杆菌噬菌体SV21(vB_PlaP_SV21)。对vB_PlaP_SV21进行了全基因组测序、比较基因组学和系统发育分析。透射电子显微镜图像显示,vB_PlaP_SV21噬菌体呈短尾病毒形态。确定对幼虫芽孢杆菌具有特异性的vB_PlaP_SV21噬菌体属于短尾病毒科。测定了该噬菌体的宿主范围和特异性、裂解量、裂解活性及形态特征。对幼虫芽孢杆菌噬菌体SV21的生物信息学分析表明,其线性44,949 bp双链DNA基因组中有77个编码序列,GC含量为39.33%。在本研究中,我们分析了所有目前已测序的幼虫芽孢杆菌噬菌体基因组,并将它们分为五个簇和一个单例。根据分子、形态和生物信息学结果,观察到在先前研究和公共数据库中被报告为单例的API480(短尾病毒)和幼虫芽孢杆菌噬菌体SV21噬菌体可共同形成一个新的簇。

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