Chen H W, Kuspa A, Keseler I M, Shimkets L J
Department of Microbiology, University of Georgia, Athens 30602.
J Bacteriol. 1991 Mar;173(6):2109-15. doi: 10.1128/jb.173.6.2109-2115.1991.
The genome of Myxococcus xanthus, which is 9,454 kbp, is one of the largest bacterial genomes. The organization of the DNA and the distribution of genes encoding social and developmental behaviors were examined by using pulsed field gel electrophoresis. Intact genomic DNA was digested with AseI into 16 restriction fragments, which were separated by contour-clamped homogeneous electric field electrophoresis, purified, and radiolabeled. Each AseI fragment was hybridized to SpeI-digested DNA and to an M. xanthus genomic library contained in yeast artificial chromosomes. Some SpeI restriction fragments and yeast artificial chromosome clones contained AseI sites and hybridized with two different AseI restriction fragments, providing evidence for the juxtaposition of these AseI restriction fragments in the chromosome. The deduced AseI physical map is circular, suggesting that this bacterium contains a single, circular chromosome. Transposable elements shown by transduction to be in or near genes of interest were located on specific AseI restriction fragments by restriction analysis and Southern hybridization. Most AseI restriction fragments contained genes involved in social and developmental behaviors.
黄色粘球菌的基因组大小为9454千碱基对,是最大的细菌基因组之一。利用脉冲场凝胶电泳技术研究了该细菌DNA的组织形式以及编码社会行为和发育行为的基因分布情况。完整的基因组DNA用AseI酶切为16个限制性片段,通过轮廓钳制均匀电场电泳进行分离,然后纯化并进行放射性标记。每个AseI片段与经SpeI酶切的DNA以及酵母人工染色体中包含的黄色粘球菌基因组文库进行杂交。一些SpeI限制性片段和酵母人工染色体克隆含有AseI位点,并与两个不同的AseI限制性片段杂交,这为这些AseI限制性片段在染色体中的并列排列提供了证据。推导的AseI物理图谱是环状的,这表明该细菌含有一条单一的环状染色体。通过转导显示存在于感兴趣基因内部或附近的转座元件,通过限制性分析和Southern杂交定位在特定的AseI限制性片段上。大多数AseI限制性片段包含参与社会行为和发育行为的基因。