Scharbaai-Vázquez Ramón, González-Caraballo Ana L, Torres-Bauzá Luis J
Department of Microbiology, San Juan Bautista School of Medicine, P.O. Box 4968, Caguas, PR 00726-4968, USA.
Plasmid. 2008 Nov;60(3):200-11. doi: 10.1016/j.plasmid.2008.07.004. Epub 2008 Sep 26.
We identified and characterized four different recombination mechanisms involved in the cointegrative transfer of the Neisseria gonorrhoeae beta-lactamase plasmid pSJ5.2 by the gonococcal 41 kb tet(M) and the Gram negative self-transmissible plasmids N3 and R64 drd-33 using an Escherichia colirecA-background. Mobilization of pSJ5.2 by the tet(M) plasmid occurred by cointegration through a replicative transposition of two IS1 elements inserted upstream from the beta-lactamase gene of pSJ5.2 and creating a IS1::beta-lactamase hybrid promoter. Two types of recombinational events occurred within the 1.8 kb BamH1-HindIII fragment of pSJ5.2 with the N3 and R64 plasmids. A non-homologous recombination was found at coordinates 1817 and 2849 of pSJ5.2 with sequences from R64. A non-homologous recombination combined with an IS26-mediated one-ended transposition was found at coordinates 1817 and 3010 of pSJ5.2 with N3. In both recombinational events, a deletion of over 1 kb of pSJ5.2 occurred. The fourth recombination event was detected in the 1.0 kb BamH1-HindIII fragment of pSJ5.2 by homologous recombination between DNA from the truncated Tn3 resolvase gene of pSJ5.2 and the resolvase sequences from R64 and N3.
我们利用大肠杆菌recA背景,鉴定并描述了四种不同的重组机制,这些机制参与了淋病奈瑟菌β-内酰胺酶质粒pSJ5.2通过淋病奈瑟菌41 kb tet(M)以及革兰氏阴性自我传递质粒N3和R64 drd-33的共整合转移。tet(M)质粒对pSJ5.2的动员是通过共整合发生的,即两个插入在pSJ5.2β-内酰胺酶基因上游的IS1元件进行复制转座,并产生一个IS1::β-内酰胺酶杂合启动子。在pSJ5.2的1.8 kb BamH1-HindIII片段内,pSJ5.2与N3和R64质粒发生了两种类型的重组事件。在pSJ5.2坐标1817和2849处,发现与R64序列发生了非同源重组。在pSJ5.2坐标1817和3010处,发现与N3发生了非同源重组并伴有IS26介导的单端转座。在这两种重组事件中,pSJ5.2均发生了超过1 kb的缺失。在pSJ5.2的1.0 kb BamH1-HindIII片段中,通过pSJ5.2截短的Tn3解离酶基因的DNA与R64和N3的解离酶序列之间的同源重组,检测到了第四种重组事件。