Mikrobiologisches Institut, Eidgenössische Technische Hochschule, ETH-Zentrum, CH-8092 Zurich, Switzerland.
Appl Environ Microbiol. 1987 Sep;53(9):2247-52. doi: 10.1128/aem.53.9.2247-2252.1987.
Spontaneous kanamycin-sensitive derivatives were obtained from Bradyrhizobium japonicum (strain 110) carrying Tn5 insertions in symbiotic gene cluster I; the derivatives were shown to have deletions of cluster I plus flanking DNA which was indicated by the absence of different copies of the repeated sequences RSalpha and RSbeta. The deletion endpoints were mapped using cloned wild-type DNA fragments containing RSalpha copies which also served as origins for overlapped cosmid cloning. The majority of the deletions resulted from recombinational fusion of two remote RSalpha copies. Novel types of repeated sequences (RSgamma, RSdelta, and RSepsilon) occurring in 12, 10, and 4 copies per genome were detected. Seven, nine, and three copies of RSgamma, RSdelta, and RSepsilon, respectively, were located near cluster I. It is concluded that the B. japonicum genome has an unusual DNA segment of >230 kilobase pairs characterized by the presence of repeated sequences and genes for symbiotic N(2) fixation.
自发获得的卡那霉素敏感衍生突变株来自于携带 Tn5 插入序列的共生基因簇 I 的日本根瘤菌(菌株 110);这些衍生突变株缺失了 I 簇及其侧翼 DNA,这表明重复序列 RSalpha 和 RSbeta 的不同拷贝不存在。缺失的终点通过使用含有 RSalpha 拷贝的克隆野生型 DNA 片段进行映射,这些拷贝也作为重叠 cosmid 克隆的起点。大多数缺失是由两个远程 RSalpha 拷贝的重组融合引起的。在基因组中分别以 12、10 和 4 个拷贝出现的新型重复序列(RSgamma、RSdelta 和 RSepsilon)被检测到。分别有 7、9 和 3 个拷贝的 RSgamma、RSdelta 和 RSepsilon 位于 I 簇附近。结论是,日本根瘤菌基因组具有一个异常的>230 千碱基对 DNA 片段,其特征是存在重复序列和共生 N(2)固定基因。