烈性噬菌体vB_VpP_AC2的分离鉴定及生物学特性分析  

Isolation and Identification of Lytic Phage vB_VpP_AC2 and Its Biological Characteristics

作　　者：黄桥栏 刘泽锟 何嘉欣 苏润斌 谢转北 黄士轩 杨美艳 HUANG Qiaolan;LIU Zekun;HE Jiaxin;SU Runbin;XIE Zhuanbei;HUANG Shixuan;YANG Meiyan(College of Food Science,South China Agricultural University,Guangdong Provincial Key Laboratory of Food Quality and Safety,Guangzhou 510642,China;College of Agriculture,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]华南农业大学食品学院,广东广州510642 [2]华南农业大学农学院,广东广州510642

出　　处：《现代食品科技》2023年第7期42-52,共11页Modern Food Science and Technology

基　　金：国家自然科学基金项目(32202194);广东省基础与应用基础研究重大项目(2020B0301030005)。

摘　　要：该研究从水产市场污水中分离纯化副溶血性弧菌噬菌体,并对其中一株噬菌体vB_VpP_AC2(AC2)进行分离鉴定、基因组及部分生物学特性分析,探究该噬菌体作为副溶血性弧菌生防制剂的潜力。噬菌体AC2的噬菌斑透明且边缘清晰,透明部分的直径约为1.12 mm,无可见晕圈。噬菌体AC2的全基因组序列长44270 bp,鸟嘌呤-胞嘧啶(GC)含量为49.30%,共预测到55个假定的开放阅读框,其中37个与编码已知功能蛋白质的基因相似(占67.27%),包括一个类似holin功能的蛋白(AC2_gp43,UTQ72417.1),该功能蛋白首次在Maculvirus属中被表征。经蛋白网络与末端酶系统发育树分析鉴定,噬菌体AC2属于自转录病毒科(Autograohiviridae),Maculvirus属,与噬菌体vB_VpaP_MGD1的ANIb值最高,为95.62%。在生物学特性方面,噬菌体AC2能够裂解30.77%的受测菌株,其最佳感染复数为0.001~0.1,一步生长曲线显示AC2的潜伏期为20 min,裂解期为35 min,裂解量为143 pfu/cell。总之,噬菌体AC2的分离鉴定不仅可为相关功能蛋白的研究提供参考,还可为水产食品安全提供较高应用价值的噬菌体资源。Vibrio parahaemolyticus phages were isolated and purified from fish market effluent.One particular phage,phage vB_VpP_AC2(AC2),was isolated,genetically sequenced,and subjected to partial biological analysis to investigate its potential as a biocontrol agent against V.parahaemolyticus.The viral plaques of phage AC2 were transparent with clear boundaries;the diameter of the transparent parts was approximately 1.12 mm,with no visible halos.The genome sequence of phage AC2 was 44270 bp long,with a guanine-cytosine content of 49.30%.In all,55 putative open reading frames were predicted,and 37(67.27%)of these were similar to genes encoding proteins of known function,including a holin-like protein(AC2_gp43,UTQ72417.1)that was first characterized in the genus Maculvirus.The results of protein-protein interaction network and terminase phylogenetic analyses showed that phage AC2 belongs to the family Autograohiviridae and genus Maculvirus.The Vibrio phage vB_VpaP_MGD1 shared the highest Average Nucleotide Identity based on BLAST(ANIb)value with AC2,at 95.62%.Biological analysis revealed that phage AC2 lysed 30.77%of the tested strains,with optimal multiplicities of infection of 0.001~0.1.Its one-step growth curve showed that its latent and lytic phases were 20 and 35 min long,respectively,with a burst size of 143 pfu/cell.In summary,the isolation and identification of phage AC2 provided useful information on its functional proteins,indicating that it is a valuable phage for aquatic food safety.

关 键 词：副溶血性弧菌 噬菌体 基因组 分离鉴定 生物学特性 

分 类 号：TS201.3[轻工技术与工程—食品科学]