Triplett Lindsay R, Zhao Youfu, Sundin George W
Department of Plant Pathology, Michigan State University, 103 Center for Integrated Plant Systems, East Lansing, MI 48824, USA.
Appl Environ Microbiol. 2006 Nov;72(11):7359-64. doi: 10.1128/AEM.01159-06. Epub 2006 Sep 8.
PCR-based subtractive hybridization was used to isolate sequences from Erwinia amylovora strain Ea110, which is pathogenic on apples and pears, that were not present in three closely related strains with differing host specificities: E. amylovora MR1, which is pathogenic only on Rubus spp.; Erwinia pyrifoliae Ep1/96, the causal agent of shoot blight of Asian pears; and Erwinia sp. strain Ejp556, the causal agent of bacterial shoot blight of pear in Japan. In total, six subtractive libraries were constructed and analyzed. Recovered sequences included type III secretion components, hypothetical membrane proteins, and ATP-binding proteins. In addition, we identified an Ea110-specific sequence with homology to a type III secretion apparatus component of the insect endosymbiont Sodalis glossinidius, as well as an Ep1/96-specific sequence with homology to the Yersinia pestis effector protein tyrosine phosphatase YopH.
基于聚合酶链反应（PCR）的消减杂交技术被用于从苹果和梨致病的解淀粉欧文氏菌菌株Ea110中分离序列，这些序列在三种具有不同宿主特异性的密切相关菌株中不存在：仅对悬钩子属植物致病的解淀粉欧文氏菌MR1；亚洲梨枝枯病的病原菌梨火疫欧文氏菌Ep1/96；以及日本梨细菌性枝枯病的病原菌欧文氏菌属菌株Ejp556。总共构建并分析了六个消减文库。回收的序列包括III型分泌成分、假定的膜蛋白和ATP结合蛋白。此外，我们鉴定出一个与昆虫内共生菌舌蝇索氏菌的III型分泌装置成分具有同源性的Ea110特异性序列，以及一个与鼠疫耶尔森菌效应蛋白酪氨酸磷酸酶YopH具有同源性的Ep1/96特异性序列。
