摘要
为控制铜绿假单胞菌的感染或污染提供生物制剂,分离到1株新的铜绿假单胞菌噬菌体,并研究其生物学特性。以铜绿假单胞菌为宿主细菌,从山西太谷某养鸡场的污水粪便混合物中分离出噬菌体。采用双层琼脂法对该噬菌体进行了分离和表征鉴定,用透射电镜(TEM)观察噬菌体的形态和大小,同时测定了噬菌体滴度、感染复数(MOI)、体外抑菌活性、一步生长曲线、温度、pH和氯仿稳定性。提取分离的噬菌体DNA,通过测序方法进行序列分析。结果显示,本研究分离出1株铜绿假单胞菌的烈性噬菌体,透射电镜显示该噬菌体属于肌病毒科,命名为vB_PaeM_TG2。噬菌斑大小为1~2mm,最佳MOI为0.1,潜伏期约为10min,暴发期约为30 min。vB_PaeM_TG2对温度、氯仿和pH具有良好的耐受性。vB_PaeM_TG2是双链环状DNA,基因组大小为206910 bp,平均GC含量为62.07%,预测有387个开放阅读框。结果表明,分离的铜绿假单胞菌噬菌体vB_PaeM_TG2潜伏期较短,裂解能力强,在高温、高氯仿浓度和宽pH范围下比较稳定。本研究为铜绿假单胞杆菌病的防治提供了新的理论支持。
This study aims to isolate a novel Pseudomonas aeruginosa(P.aeruginosa)phage,and investigate its biological characteristics.This study will provide biological agents for controlling the infection or pollution of P.aeruginosa.Using P.aeruginosa as host,we isolated phage from the sewage and feces mixture of a chicken farm at Taigu,Shanxi Province.Double-layer agar method was used to isolate and characterize the phage.Bacteriophage morphology and size were examined with a transmis-sion electron microscope(TEM).Simultaneously,the bacteriophage titer,multiplicity of infection(MOI),antibacterial activity in vitro,one-step growth curve,stability of temperature,pH and chloroform were determined.Isolated phage DNA was extracted and bioinformatics analysis by sequencing.The results showed that present study has isolated a virulent bacteriophage of P.aeruginosa.TEM showed that the phage belonged to the family Myoviridae,named vB_PaeM_TG2.The plaque size is 1—2 mm,the optimal MOI is 0.1,the incubation period is about 10 min,and the outbreak period is about 30 min.vB_PaeM_TG2 was highly tolerant to temperatures,chloroform and a wide range of pH.vB_PaeM_TG2 had double-stranded DNA with 206910 bp genome size and an average GC content of 62.07%.387 open reading frames are predicted.In conclusion that of isolated P.aeruginosa phage vB_PaeM_TG2 with short incubation period,strong lytic activity,stability under high temperatures,high chloroform concentrations and a wide range of pH.This research had provided a new theoretical support for the prevention and treatment of P.aeruginosa.
作者
刘倩
王晓蓉
苏敏
谢常红
谭姗姗
闫艺
赵宇军
任建乐
牛胜
尚桂军
梁立滨
武星辰
李俊平
王颖
田文霞
LIU Qian;WANG Xiao-rong;SU Min;XIE Chang-hong;TAN Shan-shan;YAN Yi;ZHAO Yu-jun;REN Jian-le;NIU Sheng;SHANG Gui-jun;LIANG Li-bin;WU Xing-chen;LI Jun-ping;WANG Ying;TIAN Wen-xia(College of Veterinary Medicine,Shanxi Agricultural University,Jinzhong 030801,China;Shanxi Key Laboratory of Protein Structure Determination,Shanxi Academy of Advanced Research and Innovation,Taiyuan 030032,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2023年第6期735-742,共8页
Chinese Veterinary Science
基金
国家自然科学基金项目(32202808)
山西省科技创新人才团队项目(202204051001022)
山西省现代农业产业技术体系建设专项资金(2021-07,2022-07)
山西省1331工程项目(20211331-13)
功能蛋白结构解析山西省重点实验室资助项目(202104010910006)
山西农业大学博士科研启动经费(2020BQ63)。
