Gasc A M, Kauc L, Barraillé P, Sicard M, Goodgal S
Centre de Recherche de Biochimie et de Génétique Cellulaires, Centre National de la Recherche Scientifique, Toulouse, France.
J Bacteriol. 1991 Nov;173(22):7361-7. doi: 10.1128/jb.173.22.7361-7367.1991.
A physical map of the Streptococcus (Diplococcus) pneumoniae chromosome, which is circular and 2,270 kbp in circumference, has been constructed. The restriction enzymes ApaI, SmaI, and SacII were used to digest intact chromosomes, and the fragments were resolved by field inversion gel electrophoresis (FIGE). The digests produced 22, 20, and 29 fragments, respectively. The order of the fragments was deduced from Southern blot hybridization of isolated labeled fragments to separated fragments of the various restriction digests. Genetic markers were correlated with the physical map by transformation of recipient cells with FIGE-isolated DNA fragments derived from genetically marked S. pneumoniae strains. In addition, markers were mapped by the hybridization of cloned genes to FIGE-separated restriction fragments. Six rRNA gene (rrn) clusters were mapped by hybridization to rrn-containing fragments of Haemophilus influenzae.
已构建出肺炎链球菌(双球菌)染色体的物理图谱,该染色体呈环状,周长为2270千碱基对。使用限制性内切酶ApaI、SmaI和SacII消化完整的染色体,片段通过场反转凝胶电泳(FIGE)进行分离。消化分别产生了22个、20个和29个片段。片段的顺序是通过将分离的标记片段与各种限制性消化的分离片段进行Southern印迹杂交推导出来的。通过用源自基因标记的肺炎链球菌菌株的FIGE分离的DNA片段转化受体细胞,将遗传标记与物理图谱相关联。此外,通过将克隆基因与FIGE分离的限制性片段杂交来定位标记。通过与流感嗜血杆菌含rrn的片段杂交,定位了六个rRNA基因(rrn)簇。