Kameoka D, Lezhava A, Zenitani H, Hiratsu K, Kawamoto M, Goshi K, Inada K, Shinkawa H, Kinashi H
Department of Molecular Biotechnology, Hiroshima University, 1-4-1 Kagamiyama, Higashi-Hiroshima 739-8527, Japan.
J Bacteriol. 1999 Sep;181(18):5711-7. doi: 10.1128/JB.181.18.5711-5717.1999.
A filamentous soil bacterium, Streptomyces griseus 2247, carries a 7. 8-Mb linear chromosome. We previously showed by macrorestriction analysis that mutagenic treatments easily caused deletions at both ends of its linear chromosome and changed the chromosome to a circular form. In this study, we confirmed chromosomal circularization by cloning and sequencing the junction fragments from two deletion mutants, 404-23 and N2. The junction sequences were compared with the corresponding right and left deletion end sequences in the parent strain, 2247. No homology and a 6-bp microhomology were found between the two deletion ends of the 404-23 and N2 mutants, respectively, which indicate that the chromosomal circularization was caused by illegitimate recombination without concomitant amplification. The circularized chromosomes were stably maintained in both mutants. Therefore, the chromosomal circularization might have occurred to prevent lethal deletions, which otherwise would progress into the indispensable central regions of the chromosome.
一种丝状土壤细菌,灰色链霉菌2247,携带一条7.8兆碱基对的线性染色体。我们之前通过宏观限制性分析表明,诱变处理很容易导致其线性染色体两端发生缺失,并使染色体转变为环状形式。在本研究中,我们通过克隆和测序来自两个缺失突变体404 - 23和N2的连接片段,证实了染色体环化。将连接序列与亲本菌株2247中相应的右侧和左侧缺失末端序列进行了比较。在404 - 23和N2突变体的两个缺失末端之间分别未发现同源性和6个碱基对的微同源性,这表明染色体环化是由非法重组引起的,且没有伴随扩增。环化的染色体在两个突变体中都能稳定维持。因此,染色体环化可能是为了防止致死性缺失的发生,否则这些缺失会延伸到染色体不可或缺的中心区域。