Mergeay M, Gerits J
J Bacteriol. 1978 Jul;135(1):18-28. doi: 10.1128/jb.135.1.18-28.1978.
Various F' plasmids of Escherichia coli K-12 could be transferred into mutants of the soil strain 6.2, classified herein as a Pseudomonas fluorescens biotype IV. This strain was previously found to receive Flac plasmid (N. Datta and R.W. Hedges, J. Gen Microbiol. 70:453-460, 1972). ilv, leu, met, arg, and his auxotrophs were complemented by plasmids carrying isofunctional genes; trp mutants were not complemented or were very poorly complemented. The frequency of transfer was 10(-5). Subsequent transfer into other P. fluorescens recipients was of the same order of magnitude. Some transconjugants were unable to act as donors, and these did not lose the received information if subcultured on nonselective media. Use of F' plasmids helped to discriminate metabolic blocks in P. fluorescens. In particular, metA, metB, and argH mutants were so distinguished. In addition, F131 plasmid carrying the his operon and a supD mutation could partially relieve the auxotrophy of thr, ilv, and metA13 mutants, suggesting functional expression of E. coli tRNA in P. fluorescens. In P. fluorescens metA Rifr mutants carrying the F110 plasmid, which carried the E. coli metA gene and the E. coli rifs allele, sensitivity to rifampin was found to be dominant at least temporarily over resistance. This suggests interaction of E. coli and P. fluorescens subunits of RNA polymerase. his mutations were also complemented by composite P plasmids containing the his-nif region of Klebsiella pneumoniae (plasmids FN68 and RP41). nif expression could be detected by acetylene reduction in some his+ transconjugants. The frequency of transfer of these P plasmids was 5 X 10(-4).
大肠杆菌K - 12的各种F'质粒能够转移到土壤菌株6.2的突变体中,该菌株在此被归类为荧光假单胞菌生物型IV。先前发现该菌株能接受Flac质粒(N. 达塔和R.W. 赫奇斯,《普通微生物学杂志》70:453 - 460, 1972)。ilv、leu、met、arg和his营养缺陷型由携带同功能基因的质粒互补;trp突变体未得到互补或互补效果很差。转移频率为10^(-5)。随后转移到其他荧光假单胞菌受体中的频率也在同一数量级。一些转接合子不能作为供体,并且在非选择性培养基上继代培养时不会丢失所获得的信息。使用F'质粒有助于鉴别荧光假单胞菌中的代谢阻断。特别是,metA、metB和argH突变体就是这样被区分出来的。此外,携带his操纵子和supD突变的F131质粒能够部分缓解thr、ilv和metA13突变体的营养缺陷,这表明大肠杆菌的tRNA在荧光假单胞菌中具有功能表达。在携带F110质粒(该质粒携带大肠杆菌metA基因和大肠杆菌rifs等位基因)的荧光假单胞菌metA Rifr突变体中,发现对利福平的敏感性至少在短期内对耐药性占主导。这表明大肠杆菌和荧光假单胞菌的RNA聚合酶亚基之间存在相互作用。his突变也由含有肺炎克雷伯菌his - nif区域的复合P质粒(质粒FN68和RP41)互补。在一些his +转接合子中可通过乙炔还原检测到nif表达。这些P质粒的转移频率为5×10^(-4)。