RAPID METHOD TO CHARACTERIZE LACTOCOCCAL BACTERIOPHAGE GENOMES
RAPID METHOD TO CHARACTERIZE LACTOCOCCAL BACTERIOPHAGE GENOMES
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DOI：
10.1128/aem.57.1.283-288.1991
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发表时间：
1991-01-01
影响因子：
4.4
通讯作者：
KLAENHAMMER, TR
中科院分区：
文献类型：
--
作者：
HILL, C;MASSEY, IJ;KLAENHAMMER, TR
我们提出了一种快速分离和分析噬菌体DNA的方法。细胞被感染后，噬菌体复制正常进行30 - 60分钟。在DNA包装和细胞破裂之前，收集感染细胞（1ml）并使用溶菌酶和十二烷基硫酸钠联合处理进行裂解。回收的总DNA富含噬菌体基因组，并且噬菌体DNA的限制性片段可以很容易地在琼脂糖凝胶上可视化。该方法用于粗略比较不同时期从不同奶酪植物中分离的九种乳球菌噬菌体的基因组。该方法还用于观察ptr2030诱导的流产感染对噬菌体nck202.31在其同源宿主乳酸乳球菌NCK203中的复制的抑制作用。
We present a rapid method to isolate and analyze bacteriophage DNA. Cells are infected and phage replication is allowed to proceed normally for 30 to 60 min. Prior to DNA packaging and cell bursts, the infected cells (1 ml) are harvested and lysed by using a combination of lysozyme and sodium dodecyl sulfate treatments. The total DNA recovered is enriched for phage genomes, and restriction fragments of the phage DNA can be readily visualized on agarose gels. This method was used to grossly compare the genomes of nine lactococcal phages isolated from different cheese plants at different times. The method was also used to visualize the inhibitory effects of pTR2030-induced abortive infection on the replication of phage nck202.31 in its homologous host, Lactococcus lactis NCK203.