Genetic recombination in fused spheroplasts of Providence alcalifaciens

Journal of General Microbiology
J N CoetzeeG Lecatsas

Abstract

Spheroplasts of Providence alcalifaciens strain P29 auxotrophs were prepared by combined treatment with glycine and lysozyme-EDTA. About 15% of spheroplasts had areas of cytoplasmic membrane exposed where cell wall was absent. The spheroplasts of different auxotrophs were mixed pairwise and fusion was attempted with polyethylene glycol or nascent calcium phosphate. After spheroplasts had regenerated to bacterial forms selection was made for recombinants. Recombinants arose at frequencies of 3.8 X 10(-6) to 1.7 X 10(-7) per spheroplast initially present, by both methods of fusion. The frequency was strongly dependent on the number of chromosomal loci used in selection. The possible order of five loci was determined and this corresponded to that on the closely related Proteus mirabilis chromosome. Control experiments excluded possibilities of auxotrophic reversion, conjugation, transformation, transfection or transduction as explanations of the results. Analysis of prototrophic clones yielded stable prototrophs or mixtures of stable prototrophs and stable recombinants. Parental types were not encountered. Unselected markers segregated among recombinants. It was concluded that the formation of recombinant bacteria was due to spher...Continue Reading

Citations

Jun 23, 2009·Current Microbiology·Bheong-Uk LeeKye-Heon Oh
Oct 20, 2011·Current Genetics·Matej Vesteg, Juraj Krajčovič
Oct 1, 1993·Journal of Biotechnology·J O de MoraisC E Lopes
Sep 2, 2003·Microbiology·Jean-Pierre Gratia, Marc Thiry
Sep 1, 1985·Applied and Environmental Microbiology·M E BarrosD R Woods
Apr 1, 1983·Journal of Bacteriology·M L Stahl, P A Pattee
Jan 1, 1981·Journal of Bacteriology·F GötzM Lindberg

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