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丁香假单胞菌菜豆致病变种八个切除质粒的特性分析

Characterization of eight excision plasmids of Pseudomonas syringae pv. phaseolicola.

作者信息

Szabo L J, Mills D

出版信息

Mol Gen Genet. 1984;195(1-2):90-5. doi: 10.1007/BF00332729.

Abstract

Pseudomonas syringae pv. phaseolicola strain LR719 contains a 150 kilobase pair (kb) plasmid pMC7105, stably integrated into its chromosome. Occasionally, single colony isolates of this strain contain an excision plasmid. Eight unique excision plasmids were selected and characterized by BamHI restriction endonuclease and blot hybridization analyses. These plasmids ranged in size from 35 to 270 kb; the largest contained approximately 130 kb of chromosomal DNA sequences. Restriction maps of pMC7105 were developed to deduce the site of integration and to identify the fragments in which recombination occurred to produce each excision plasmid. The eight excision plasmids were arranged into five classes based on the sites where excision occurs. A 20 kb region of pMC7105, which includes BamHI fragment 9 and portions of adjacent fragments, is present in all excision plasmids and thought to contain the origin of replication. The site of integration on pMC7105 maps within BamHI fragment 8. This fragment shows homology with seven other BamHI fragments of pMC7105 and with five chromosomal fragments identified among the excision plasmids. The data strongly suggest that the integration of pMC7105 may have occurred at a repetitive sequence present on the chromosome and on the plasmid.

摘要

菜豆丁香假单胞菌菜豆致病变种LR719菌株含有一个稳定整合到其染色体中的150千碱基对(kb)的质粒pMC7105。偶尔,该菌株的单菌落分离物会含有一个切除质粒。选择了八个独特的切除质粒,并通过BamHI限制性内切酶和印迹杂交分析对其进行了表征。这些质粒的大小在35至270 kb之间;最大的质粒包含约130 kb的染色体DNA序列。构建了pMC7105的限制性图谱,以推断整合位点,并鉴定发生重组以产生每个切除质粒的片段。根据切除发生的位点,将八个切除质粒分为五类。所有切除质粒中都存在pMC7105的一个20 kb区域,该区域包括BamHI片段9和相邻片段的部分,并且被认为包含复制起点。pMC7105上的整合位点位于BamHI片段8内。该片段与pMC7105的其他七个BamHI片段以及在切除质粒中鉴定出的五个染色体片段具有同源性。数据强烈表明,pMC7105的整合可能发生在染色体和质粒上存在的重复序列处。

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