源于克雷伯氏菌Klebsiellasp.AC-11的3株噬菌体的生物学特性
Biological characteristics of 3 lytic bacteriophages from the same host Klebsiella sp. AC-11
-
摘要: 从国家海洋局第三海洋研究所海水养殖池中分离得到一株细菌AC-11,16SrDNA序列分析表明其与肺炎克雷伯氏菌肺炎亚种(Klebsiella pneumoniae subsp. pneumoniae)具有最高的序列同源性达99.78%。以此菌株为宿主菌,于2008年11月采用双层平板法分离得到3株裂解性噬菌体KP1,KP2和KP3,其中KP2分离自同一海水养殖池,另外2株则来源于厦门筼筜湖海水样品。分别对宿主菌及噬菌体进行了电镜形态观察,并对噬菌体进行了BglⅡ和EcoRⅤ两种酶的限制性酶切分析,发现它们均属于长尾噬菌体科(Siphoviridae),但具有不同的酶切图谱;同时测定了3株噬菌体的最佳感染复数、一步生长曲线及对温度、pH、紫外线、乙醚和氯仿等理化因素的敏感性等生理特性。实验结果表明3株噬菌体具有不同的生物学特性,为不同株型。本研究丰富了对噬菌体多样性的认识,并为进一步应用噬菌体诊断和治疗潜在的由肺炎克雷伯氏菌引发的疾病及细菌分型等研究提供了理论基础。Abstract: The bacterial strain AC-11 was isolated from marine aquaculture pond of the Third Institute of Oceanography in Xiamen, China, and its 16SrDNA sequence demonstrated 99.78% similarity to Klebsiella pneumoniae subsp. pneumoniae. A total of 3 lytic phages KP1, KP2 and KP3 were isolated with this bacterial strain as the host using double-layer agar method. Phage KP2 was from the same aquaculture pond where the host strain was isolated, and the other two phages were from Yuan Dang Lake (marine water) in Xiamen, China. Morphology of both phages and host were observed under transmission electron microscope. The restriction patterns of nucleic acid of the phages were estimated by dsDNA restriction enzyme BglⅡand EcoRⅤ. All of the 3 phages belonged to family Siphoviridae, but with different genome fingerprint patterns. The physiological characteristics of the phages such as the optimal multiplicity of infection, the one-step growth curves and the sensitivity to temperature, pH, UV light, ether and chloroform were also tested. The results indicated that the phages had different biological characteristics and thus belonged to different strain types. This could contribute to the diversity of phages, the diagnosis and treatment of potential diseases caused by Klebsiella pneumoniae and typing of the relating Klebsiella bacteria.
-
兰琳, 林锡煌, 彭益强, 等. 耐高浓度甘油的1,3-丙二醇生产菌的诱变选育[J]. 华侨大学学报, 2007, 28(4): 399-402. 杜军, 纪晓俊, 胡南, 等. 耐高糖高产2,3-丁二醇产酸克雷伯氏杆菌的选育[J]. 中国生物工程杂志, 2008, 28(6 ):89-92. 邬小兵, 徐惠娟, 丛一博, 等. 产酸克雷伯氏菌耐氧产氢及其可溶性氢酶耐氧特性研究[J]. 生物技术, 2008, 18(4):50-51. PODSCHUN R, ULLMANN U. Klebsiella spp. as Nosocomial Pathogens:Epidemiology,Taxonomy, Typing Methods, and Pathogenicity Factors [J]. Clinical Microbiology Reviews, 1998, 11(4):589-603. BRISSE S, DUIJKEREN E. Identification and antimicrobial susceptibility of 100 Klebsiella animal clinical isolates [J]. Veterinary Microbiology, 2005, 105:307-312. 何礼洋, 韩文瑜, 雷连成, 等. 肺炎克雷伯氏菌Ⅲ型菌毛的研究进展[J]. 中国兽医杂志, 2007, 43(12):50-51. 王林峰, 王选锭. 肺炎克雷伯氏菌耐药机制研究进展[J]. 中国抗生素杂志, 2004, 29(6): 324-328. STRUVE C, KROGFELT A K. Pathogenic potential of environmental Klebsiella pneumoniae isolates [J]. Environmental Microbiology, 2004, 6(6):584-590. 邓国成, 罗霞, 江小燕, 等. 鳗肺炎克雷伯菌的分离与鉴定[J]. 上海海洋大学学报, 2009, 18(2):193-197. 黄建军, 胡晓敏, 饶贤才, 等. 铜绿假单胞菌噬菌体PaP2生物学特性的研究[J]. 第三军医大学学报, 2004 ,26(13):1133-1136. 刘明河. 细菌病毒--噬菌体的应用概况[J]. 微生物学杂志, 2002, 22(4):56-57. SKURNIK M, STRAUCH E. Phage therapy: Facts and fiction [J]. International Journal of Medical Microbiology, 2006, 296:5-14. BORYSOWSKI J, GORSKI A. Is phage therapy acceptable in the immunocompromised host? [J]. International Journal of Infectious Diseases, 2008, 12:466-471. NAKAI T, PARK S C. Bacteriophage therapy of infectious diseases in aquaculture [J]. Research in Microbiology, 2002, 153:13-18. ADAMS M H. Bacteriophages[M]. Interscience Publishers, Inc., New York, 1959: 592. 萨姆布鲁克 J,拉塞尔 D W.分子克隆实验指南[M].第3版.黄培堂等译.北京:科学出版社, 2002:170-243. 奥斯伯 F M, 布伦特 R, 金斯顿 E, 等. 精编分子生物学实验指南[M].第5版.金由辛,包慧中,赵丽云等译.北京:科学出版社, 2008: 39. 漆辉州, 陈红, 敖敬群, 等 一株深海中等嗜热嗜酸菌的分离及鉴定[J]. 海洋学报, 2009, 31(2):152-158. JIANG S C, KELLOGG C A, PAUL J H. Characterization of marine temperate phage-host systems isolated from Mamala Bay, Oahu, Hawaii [J]. Applied and Environmental Microbiology, 1998, 64(2):535-542. GARNEAU J E, TREMBLAY D M, MOINEAU S. Characterization of 1706, a virulent phage from Lactococcus lactis with similarities to prophages from other Firmicutes [J]. Virology, 2008, 373:298-309. 宁淑香, 聂丽平, 陆敏, 等. VP10 -一株海洋噬菌体的分离及特性研究[J]. 水产科学 2000, 19(2):14-16. 邱德全, 蔺红苹, 谭龙艳. 一株副溶血弧菌噬菌体生理特性的研究[J]. 微生物学通报, 2007, 34(4):735-739. 孙佳, 叶德赞, Kochel A, 等. 裂解性弧菌噬菌体的分离及生理特性[J]. 微生物学报, 2008, 48(6):780-784. ACKERMANN H W. 5500 Phages examined in the electron microscope [J]. Archives of Virology, 2007, 152: 227-243. 于龙, 温占波, 杨文慧, 等. 一株粘质沙雷氏菌烈性噬菌体污水分离及特性[J]. 微生物学报, 2008, 48(4): 498-502. 杨水云, 郭春林, 赵文明. 理化因素对苏云金芽孢杆菌噬菌体活性影响的研究[J]. 陕西师范大学学报, 1998, 26:225-228. 凌体淑, 叶迎春, 赵世鸿, 等. 细菌荚膜染色的创新[J]. 现代检验医学杂志, 2004, 19(4):35-36.
点击查看大图
计量
- 文章访问数: 1705
- HTML全文浏览量: 16
- PDF下载量: 1035
- 被引次数: 0