Interdomain conjugal transfer of DNA from bacteria to archaea - PubMed (original) (raw)

Interdomain conjugal transfer of DNA from bacteria to archaea

Jeremy A Dodsworth et al. Appl Environ Microbiol. 2010 Aug.

Abstract

Escherichia coli transforms the methanogenic archaeon Methanococcus maripaludis at frequencies ranging from 0.2 x 10(-6) to 2 x 10(-6) per recipient cell. Transformation requires cell-to-cell contact, oriT, and tra functions, is insensitive to DNase I, and otherwise displays hallmarks of conjugation.

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Figures

FIG. 1.

Genetic analysis of M. maripaludis transformants. (A) A schematic diagram of the nrpR gene and flanking region in the M. maripaludis genome and the nrpR::pac region of the gene replacement constructs pTAP1, pTAP4, and pTAP6, harboring mob-oriT-rep, mob-oriT, and RP4-oriT, respectively (open boxes). Primers for PCR analyses are shown with arrowheads, and the probe for Southern analysis is indicated. gDNA, genomic DNA. (B) Southern blot and PCR analyses of DNA extracted from putative pTAP1, pTAP4, and pTAP6 transformants of M. maripaludis. Arrows indicate the signature band (6.3 kb) for double crossover (c/o), 5′ crossover, and 3′ crossover. Positive and negative PCR amplifications are shown as “+” and “−,” respectively. WT, wild-type M. maripaludis S2; MM500, nrpR deletion mutant generated by PEG-mediated transformation with an nrpR::_pac_-containing construct (10).

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