Widjaja R, Suwanto A, Tjahjono B
Department of Biology, Faculty of Science and Mathematics, Bogor Agricultural University, Indonesia.
FEMS Microbiol Lett. 1999 Jun 1;175(1):59-68. doi: 10.1111/j.1574-6968.1999.tb13602.x.
Xanthomonas campestris is an important plant pathogenic bacterium which causes severe diseases in a wide variety of plant species. We have generated a macrorestriction map of the X. campestris (axonopodis) pv. glycines chromosome employing pulsed-field gel electrophoresis (PFGE). Restriction endonucleases PacI (5'-TTAATTAA), PmeI (5'-GTTTAAAC) and SwaI (5'-ATTTAAAT) digested the chromosomal DNA into three, five, and five fragments, respectively. In addition, intron-encoded restriction endonuclease I-CeuI was employed to locate the position of the 23S rRNA genes (rrlA and rrlB). All of the generated restriction fragments were aligned along the chromosome using multiple restriction enzyme digestion and two-dimensional PFGE (2-D PFGE) in conjunction with Southern hybridization analysis. This physical map construction has revealed a single circular chromosome with a size of approximately 5 Mb. Two rRNA genes were localized on the chromosome map. Several genes involved in pathogenesis (xpsD, opsX, and pat) as well as genes involved in the biosynthesis of xanthan gum (xanAB, rfbCDAB) were also localized.
野油菜黄单胞菌是一种重要的植物病原菌,可在多种植物物种中引发严重病害。我们利用脉冲场凝胶电泳(PFGE)构建了野油菜黄单胞菌(致病变种)大豆致病变种的染色体宏观限制性图谱。限制性内切酶PacI(5'-TTAATTAA)、PmeI(5'-GTTTAAAC)和SwaI(5'-ATTTAAAT)分别将染色体DNA切割成三个、五个和五个片段。此外,利用内含子编码的限制性内切酶I-CeuI来定位23S rRNA基因(rrlA和rrlB)的位置。通过多重酶切和二维PFGE(2-D PFGE)结合Southern杂交分析,将所有产生的限制性片段沿染色体进行比对。该物理图谱构建显示出一条大小约为5 Mb的单一环状染色体。两个rRNA基因定位在染色体图谱上。一些与致病相关的基因(xpsD、opsX和pat)以及与黄原胶生物合成相关的基因(xanAB、rfbCDAB)也被定位。