IncP plasmids are unusually effective in mediating conjugation of Escherichia coli and Saccharomyces cerevisiae:: Involvement of the Tra2 mating system
IncP plasmids are unusually effective in mediating conjugation of Escherichia coli and Saccharomyces cerevisiae:: Involvement of the Tra2 mating system
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DOI：
10.1128/jb.180.24.6538-6543.1998
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发表时间：
1998-12-01
影响因子：
3.2
通讯作者：
Wilkins, BM
中科院分区：
文献类型：
--
作者：
Bates, S;Cashmore, AM;Wilkins, BM
构建了含有质粒F(IncFI)、ColIb-P9(IncI1)和Rp4/RP1(IncPα)的可移动穿梭质粒，以测试同源接合系统介导基因从大肠杆菌向酿酒酵母转移的能力。只有Pα系统对酵母产生了可检测到的动员，在30分钟内每个受体细胞的接合子达到了5×10(-5)的峰值。穿梭质粒的转移需要在顺式中携带ORIT，并在反式中提供PαTra1和Tra2核心区。Tra1核心外的基因不能提高动员效率。所有10个Tra2核心基因(trbB、-C、-D、-E、-F、-G、-H、-I、-J和-L)都需要转移到酵母中。为了评估交配对形成(MPF)系统或Pα接合系统的DNA处理装置是否在跨王国转移中起关键作用，设计了一种使用INQ为基础的穿梭质粒指定其自己的DNA处理系统的分析方法。RP1而不是ColIb将构建物动员到酵母上，表明由Tra2核心基因和Traf决定的MPF复合体是Pα系统在介导基因从细菌到真核生物转移中显著肥沃的主要原因。
Mobilizable shuttle plasmids containing the origin-of-transfer (oriT) region of plasmids F (IncFI), ColIb-P9 (IncI1), and RP4/RP1 (IncP alpha) were constructed to test the ability of the cognate conjugation system to mediate gene transfer from Escherichia coli to Saccharomyces cerevisiae. Only the P alpha system caused detectable mobilization to yeast, giving peak values of 5 x 10(-5) transconjugants per recipient cell in 30 min. Transfer of the shuttle plasmid required carriage of oriT in cis and the provision in trans of the P alpha Tra1 core and Tra2 core regions. Genes outside the Tra1 core did not increase the mobilization efficiency. All 10 Tra2 core genes (trbB, -C, -D, -E, -F, -G, -H, -I, -J, and -L) required for plasmid transfer to E. coli K-12 were needed for transfer to yeast. To assess whether the mating-pair formation (Mpf) system or DNA-processing apparatus of the P alpha conjugation system is critical in transkingdom transfer, an assay using an IncQ-based shuttle plasmid specifying its own DNA-processing system nas devised. RP1 but not ColIb mobilized the construct to yeast, indicating that the Mpf complex determined by the Tra2 core genes plus traF is primarily responsible for the remarkable fertility of the P alpha system in mediating gene transfer from bacteria to eukaryotes.