Lunde M, Blatny J M, Kaper F, Nes I F, Lillehaug D
Laboratory of Microbial Gene Technology, Department of Chemistry and Biotechnology, Agricultural University of Norway, P.O. Box 5051, N-1432, Aas, Norway.
FEMS Microbiol Lett. 2000 Nov 1;192(1):119-24. doi: 10.1111/j.1574-6968.2000.tb09369.x.
We present here a new and general approach for monitoring the life cycles of temperate bacteriophages which establish lysogeny by inserting their genomes site-specifically into the bacterial host chromosome. The method is based on quantitative amplification of specific DNA sites involved in various cut-and-join events during the life cycles of the phages (i.e. the cos, attP, attB, attL and attR sites) with the use of sequence-specific primers. By comparing the amounts of these specific DNA sites at different intervals, we were able to follow the development of the lytic and lysogenic life cycles of the temperate lactococcal bacteriophage phiLC3 after infection of its bacterial host Lactococcus lactis ssp. cremoris IMN-C18.
我们在此展示一种全新的通用方法，用于监测通过将基因组位点特异性插入细菌宿主染色体而建立溶原性的温和噬菌体的生命周期。该方法基于使用序列特异性引物对噬菌体生命周期中各种切割和连接事件所涉及的特定DNA位点（即粘性末端位点、噬菌体附着位点、细菌附着位点、左向附着位点和右向附着位点）进行定量扩增。通过比较不同时间间隔这些特定DNA位点的数量，我们能够追踪温和乳球菌噬菌体phiLC3感染其细菌宿主乳酸乳球菌亚种cremoris IMN-C18后裂解和溶原性生命周期的进展。
