Song Eun-Sung, Park Young-Jin, Chae Soo-Cheon, Kim Jeong-Gu, Cho Hee-Jung, Lee Gil-Bok, Lee Byoung-Moo
National Institute of Agricultural Biotechnology, Rural Development Administration, Suwon 441-707, Korea.
Biotechnol Lett. 2006 Jul;28(13):969-77. doi: 10.1007/s10529-006-9027-5. Epub 2006 Jun 24.
Pseudomonas syringae pv. tagetis causes apical chlorosis of several plant species in the Asteraceae, including marigold. As a means to facilitate the isolation of pathogenicity genes and to characterize the genome of this bacterium, we have constructed a bacterial artificial chromosome library of P. syringae pv. tagetis strain LMG5090. The library consists of 1,536 clones with insert size ranging from 30 to 160 kb and an average size of 86 kb. Based upon colony hybridization, the BAC clone 420E23 containing the hrp/hrc gene cluster encoding the type III secretion system was identified from this library and subsequently shotgun sequenced. The hrp/hrc gene cluster of P. syringae pv. tagetis has a 23 kb sequence which contains 27 open reading frames. Comparative analysis of the hrp/hrc gene cluster of P. syringae pv. tagetis LMG5090, P. syringae pv. tomato DC3000, P. syringae pv. syringae B728a, and P. syringae pv. phaseolicola 1448A revealed that the entire hrp/hrc gene cluster of P. syringae pv. tagetis is conserved and identically arranged in all four pathovars.
丁香假单胞菌丁香致病变种可导致包括万寿菊在内的菊科多种植物出现顶端黄化现象。为便于分离致病基因并对该细菌的基因组进行表征，我们构建了丁香假单胞菌丁香致病变种菌株LMG5090的细菌人工染色体文库。该文库由1536个克隆组成，插入片段大小在30至160 kb之间，平均大小为86 kb。基于菌落杂交，从该文库中鉴定出包含编码III型分泌系统的hrp/hrc基因簇的BAC克隆420E23，并随后进行了鸟枪法测序。丁香假单胞菌丁香致病变种的hrp/hrc基因簇有一个23 kb的序列，包含27个开放阅读框。对丁香假单胞菌丁香致病变种LMG5090、丁香假单胞菌番茄致病变种DC3000、丁香假单胞菌丁香致病变种B728a和菜豆假单胞菌1448A的hrp/hrc基因簇进行比较分析，结果表明丁香假单胞菌丁香致病变种的整个hrp/hrc基因簇在所有四个致病变种中都是保守的且排列相同。
