Haines Anthony S, Akhtar Parveen, Stephens Elton R, Jones Karen, Thomas Christopher M, Perkins Caroline D, Williams Jacqueline R, Day Martin J, Fry John C
School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
Cardiff School of Biosciences, Cardiff University, PO Box 915, Cardiff CF1 3TL, UK.
Microbiology (Reading). 2006 Sep;152(Pt 9):2689-2701. doi: 10.1099/mic.0.28941-0.
Nine mercury-resistance plasmids isolated from river epilithon were assessed for their ability to retrotransfer the non-conjugative IncQ plasmid, R300B, derivatives of which have commercial uses that may result in accidental or deliberate release into the environment. Retrotransfer frequencies ranging from 2.1 x 10(-4) to 1.75 x 10(-5) were obtained for five of the nine plasmids--the remaining plasmids showed low or undetectable retrotransfer ability. The majority of the retrotransfer-proficient plasmids could not be classified by the tests used. Classical incompatibility testing with RP4 identified pQKH6, pQKH54 and pQM719 as IncP-1. Hybridization to replicon probes confirmed this for pQKH6 and pQM719 and added pQKH33. PCR with primers designed to amplify trfA and korA regions of IncP-1 plasmids did not identify any other plasmids. Plasmids pQKH6 and pQM719 but not pQKH54 produced similar SphI restriction profiles to the IncP-1beta subgroup. The complete nucleotide sequence of pQKH54 was determined, revealing it to have a complete IncP-1 backbone but belonging to a new distinct subgroup which was designated IncP-1gamma. The results emphasize the ubiquity and diversity of IncP-1 plasmids in the environment but demonstrate that plasmids of as yet unknown groups are also able to retrotransfer IncQ plasmids efficiently.
对从河流附生生物中分离出的9种耐汞质粒进行了评估,以检测它们回传非接合型IncQ质粒R300B的能力,R300B的衍生物具有商业用途,可能会意外或故意释放到环境中。9种质粒中有5种的回传频率在2.1×10⁻⁴至1.75×10⁻⁵之间,其余质粒的回传能力较低或无法检测到。大多数具有回传能力的质粒无法通过所用测试进行分类。用RP4进行的经典不相容性测试确定pQKH6、pQKH54和pQM719为IncP-1。与复制子探针杂交证实pQKH6和pQM719属于IncP-1,并新增了pQKH33。用设计用于扩增IncP-1质粒trfA和korA区域的引物进行PCR,未鉴定出任何其他质粒。质粒pQKH6和pQM719而非pQKH54产生了与IncP-1β亚组相似的SphI限制性图谱。测定了pQKH54的完整核苷酸序列,发现它具有完整的IncP-1骨架,但属于一个新的独特亚组,被命名为IncP-1γ。结果强调了IncP-1质粒在环境中的普遍性和多样性,但表明尚未知的质粒组也能够有效地回传IncQ质粒。