Genome Mapping of Plant Pathogenic Bacteria

植物病原细菌基因组图谱

基本信息

批准号：
06660047
负责人：
TAKIKAWA Yuichi
金额：
$ 1.34万
依托单位：
Shizuoka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06660047/
关键词：
Plant pathogenic bacteria Pseudomonas Gene library

项目摘要

Construction of gene libraries from several Pseudomonas plant pathogens were challenged. During these two years, the libraries were successfully constructed from P.syringae pv.phaseolicola, P.s.glycinea, P.s.maculicola, P.s.tabaci, P.s.syringae, and P.s.actinidiae. From the libraries, the clones which have homology with hrp genes and phaseolotoxin production genes were selected by hybridization and genomic walking. As for hrpregions, a map was obtained on a long area covering more than 120kb. The homologous regions in the clones from different bacteria are preserved for more than several 10kb in length in each directions beyond the known hrp region. The homologies sharply declined toward the out side of the region. As for phaseolotoxin production genes, clones were obtained from P.s.glycinea which showed homology with the borders of the phaseolotoxin production genes. The clones were aligned and mapped. The result showed that the borders from P.s.glycinea are intervened by a 12kb unknown fragment which has no homology with phaseolotoxin production genes. That strongly suggested that the phaseolotoxin production genes have moved horizontally among bacteria. Unfortunately, entire mapping using puls-field electrophoresis and PCR was unsuccessful, though, the results obtained were valuable indicating that the strategy of genome mapping can lead to the elucidation of the evolutionary process in plant pathogenic bacteria.

挑战了几种假单胞菌病原体的基因库。在这两年中，图书馆成功地从P.Syringae Pv.phaseolicola，P.S.Glycinea，P.S. -Maculicola，P.S.Tabaci，P.S.Syringae和P.S.S.Stinidiae构建。从图书馆中，通过杂交和基因组行走选择了与HRP基因和生物毒素生产基因同源的克隆。至于HRPRIGIONS，在覆盖超过120kb的长区域上获得了地图。来自不同细菌的克隆中的同源区域可在已知的HRP区域以外的每个方向上保留多个10KB的长度。同源物朝着该地区的外侧急剧下降。至于阶段性毒素的产生基因，从p.s.- glycinea获得了克隆，该基因与阶段溶毒素生产基因的边界表现出同源性。克隆对齐并映射。结果表明，来自P.S. glycinea的边界是由12KB未知片段干预，该片段与阶段性毒素生产基因没有同源性。这强烈表明，循环毒素的产生基因在细菌之间水平移动。不幸的是，使用脉冲场电泳和PCR的整个映射没有成功，获得的结果非常有价值，这表明基因组映射的策略可以导致植物致病细菌中进化过程的阐明。