Tseng Y H, Choy K T, Hung C H, Lin N T, Liu J Y, Lou C H, Yang B Y, Wen F S, Weng S F, Wu J R
Institute of Molecular Biology and Department of Botany, National Chung Hsing University, Taichung 402, Taiwan.
J Bacteriol. 1999 Jan;181(1):117-25. doi: 10.1128/JB.181.1.117-125.1999.
No plasmid was detected in Xanthomonas campestris pv. campestris 17, a strain of the causative agent of black rot in cruciferous plants isolated in Taiwan. Its chromosome was cut by PacI, PmeI, and SwaI into five, two, and six fragments, respectively, and a size of 4.8 Mb was estimated by summing the fragment lengths in these digests. Based on the data obtained from partial digestion and Southern hybridization using probes common to pairs of the overlapping fragments or prepared from linking fragments, a circular physical map bearing the PacI, PmeI, and SwaI sites was constructed for the X. campestris pv. campestris 17 chromosome. Locations of eight eps loci involved in exopolysaccharide (xanthan gum) synthesis, two rrn operons each possessing an unique I-CeuI site, one pig cluster required for yellow pigmentation, and nine auxotrophic markers were determined, using mutants isolated by mutagenesis with Tn5(pfm)CmKm. This transposon contains a polylinker with sites for several rare-cutting restriction endonucleases located between the chloramphenicol resistance and kanamycin resistance (Kmr) genes, which upon insertion introduced additional sites into the chromosome. The recA and tdh genes, with known sequences, were mapped by tagging with the polylinker-Kmr segment from Tn5(pfm)CmKm. This is the first map for X. campestris and would be useful for genetic studies of this and related Xanthomonas species.
在野油菜黄单胞菌野油菜致病变种17中未检测到质粒,该菌株是从台湾分离得到的十字花科植物黑腐病病原体。其染色体分别被PacI、PmeI和SwaI切割成5个、2个和6个片段,通过将这些酶切片段的长度相加估计其大小为4.8 Mb。基于部分酶切和Southern杂交的数据,使用重叠片段对的共同探针或连接片段制备的探针,构建了野油菜黄单胞菌野油菜致病变种17染色体的带有PacI、PmeI和SwaI位点的环状物理图谱。利用通过Tn5(pfm)CmKm诱变分离得到的突变体,确定了参与胞外多糖(黄原胶)合成的8个eps基因座、每个都具有独特I-CeuI位点的2个rrn操纵子、黄色色素形成所需的1个pig簇以及9个营养缺陷型标记的位置。该转座子包含一个多克隆位点,在氯霉素抗性基因和卡那霉素抗性(Kmr)基因之间有几个稀有切割限制内切酶的位点,插入时会在染色体中引入额外的位点。通过用来自Tn5(pfm)CmKm的多克隆位点-Kmr片段进行标记,对已知序列的recA和tdh基因进行了定位。这是野油菜黄单胞菌的第一张图谱,将有助于对该菌及相关黄单胞菌属物种进行遗传学研究。