Luzhetskyy A, Fedoryshyn M, Gromyko O, Ostash B, Rebets Y, Bechthold A, Fedorenko V
Albert-Ludwigs-University of Freiburg, Pharmazeutische Biologie, Freiburg D-79104, Germany.
Genetika. 2006 May;42(5):595-601.
Mobilizable shuttle plasmids containing the origin of transfer (oriT) region of plasmid F (IncFI), ColIb-P9 (IncI1), and RP4/RP1 (IncPalpha) were constructed to test the ability of the cognate conjugation system to mediate gene transfer from Escherichia coli to Streptomyces. The conjugative system of the IncPalpha plasmids was shown to be most effective in conjugative transfer, giving peak values of (2.7 +/- 0.2) x 10(-2) S. lividans TK24 exconjugants per recipient cell. To assess whether the mating-pair formation system or the DNA-processing apparatus of the IncPalpha plasmids is crucial in conjugative transfer, an assay with an IncQ-based mobilizable plasmid (RSF1010) specifying its own DNA-processing system was developed. Only the IncPalpha plasmid mobilized the construct to S. lividans indicating that the mating-pair formation system is primarly responsible for the promiscuous transfer of the plasmids between E. coli and Streptomyces. Dynamic of conjugative transfer from E. coli to S. lividans was investigated and exconjugants starting from the first hour of mating were obtained.
构建了含有质粒F（IncFI）、ColIb - P9（IncI1）和RP4/RP1（IncPα）转移起始区（oriT）的可动员穿梭质粒，以测试同源接合系统介导基因从大肠杆菌转移至链霉菌的能力。结果表明，IncPα质粒的接合系统在接合转移中最为有效，每个受体细胞产生的淡紫灰链霉菌TK24接合子峰值为(2.7±0.2)×10⁻²。为评估IncPα质粒的配对形成系统或DNA加工装置在接合转移中是否至关重要，开发了一种基于IncQ的可动员质粒（RSF1010）的检测方法，该质粒指定了其自身的DNA加工系统。只有IncPα质粒能将构建体转移至淡紫灰链霉菌，这表明配对形成系统主要负责质粒在大肠杆菌和链霉菌之间的广泛转移。研究了从大肠杆菌到淡紫灰链霉菌的接合转移动态，并获得了从交配后第一小时开始的接合子。
