Departments of Biochemistry and Plant Pathology, North Dakota State University, Fargo, North Dakota 58105.
Appl Environ Microbiol. 1987 Oct;53(10):2476-81. doi: 10.1128/aem.53.10.2476-2481.1987.
Restriction fragments of pCS1, a 50.6-kilobase (kb) plasmid present in many strains of Clavibacter michiganense subsp. sepedonicum ("Corynebacterium sepedonicum"), have been cloned in an M13mp11 phage vector. Radiolabeled forms of these cloned fragments have been used as Southern hybridization probes for the presence of plasmid sequences in chromosomal DNA of this organism. These studies have shown that all tested strains lacking the covalently closed circular form of pCS1 contain the plasmid in integrated form. In each case the site of integration exists on a single plasmid restriction fragment with a size of 5.1 kb. Southern hybridizations with these probes have also revealed the existence of a major repeated sequence in C. michiganense subsp. sepedonicum. Hybridizations of chromosomal DNA with deletion subclones of a 2.9-kb plasmid fragment containing the repeated sequence indicate that the size of the repeated sequence is approximately 1.3 kb. One of the copies of the repeated sequence is on the plasmid fragment containing the site of integration.
pCS1 是一种存在于许多密歇根棒杆菌亚种(“棒杆菌属”)菌株中的 50.6 千碱基(kb)质粒,其限制片段已被克隆到 M13mp11 噬菌体载体中。这些克隆片段的放射性标记形式已被用作 Southern 杂交探针,以检测该生物体染色体 DNA 中质粒序列的存在。这些研究表明,所有测试的缺乏 pCS1 共价闭合环状形式的菌株都以整合形式存在质粒。在每种情况下,整合位点存在于一个大小为 5.1 kb 的单个质粒限制片段上。这些探针的 Southern 杂交还揭示了密歇根棒杆菌亚种中存在一个主要的重复序列。用包含重复序列的 2.9 kb 质粒片段的缺失亚克隆与染色体 DNA 的杂交表明,重复序列的大小约为 1.3 kb。重复序列的一个拷贝位于包含整合位点的质粒片段上。