Lezhava A, Kameoka D, Sugino H, Goshi K, Shinkawa H, Nimi O, Horinouchi S, Beppu T, Kinashi H
Department of Fermentation Technology, Hiroshima University, Higashi-Hiroshima, Japan.
Mol Gen Genet. 1997 Jan 27;253(4):478-83. doi: 10.1007/s004380050346.
We have recently constructed a physical map of the Streptomyces griseus 2247 genome using the restriction enzymes AseI and DraI, which revealed that this strain carries a 7.8 Mb linear chromosome. Based on this map, precise macrorestriction fragment and cosmid maps were constructed for both ends of the chromosome, which localized the afsA gene 150 Kb from the left end. Two afsA- mutants were found to have suffered chromosomal deletions that removed the afsA locus. The sizes of the deletions were 20 and 130 Kb at the right end and 180 and 350 kb at the left end, respectively. Hybridization experiments using cosmids carrying a deletion endpoint indicated that the ends of the chromosome in the mutants were fused to form a circular chromosome.
我们最近利用限制性内切酶AseI和DraI构建了灰色链霉菌2247基因组的物理图谱,结果显示该菌株携带一条7.8 Mb的线性染色体。基于此图谱,构建了染色体两端精确的宏观限制性片段图谱和黏粒图谱,将afsA基因定位在距左端150 Kb处。发现两个afsA-突变体发生了染色体缺失,缺失了afsA基因座。缺失大小在右端分别为20和130 Kb,在左端分别为180和350 kb。使用携带缺失端点的黏粒进行的杂交实验表明,突变体中染色体的两端融合形成了环状染色体。