SUMMARY: Spheroplasts were prepared from two carotenoid pigment mutants of named NgR and A which were obtained by the chemical mutagenesis of the wild type strain A with -methyl--nitro--nitrosoguanidine. The carotenoid pigments and the α- and β-mycolic acids were taken as genetic markers and the recombinants were selected on the basis of their colour on Löwenstein-Jensen medium. Spheroplasts of the two mutants were mixed in a 1:1 ratio and were treated with 40% (w/v) polyethylene glycol 6000 for 5 min at 37 °C. The frequency of NgR × A recombination in optimal conditions was about 2.5 × 10. The recombinants selected on the basis of their carotenoid pigment profile were also tested for their α- and β-mycolic acids as a second genetic marker. The results were further confirmed by electron microscopy. The optimal conditions, for spheroplast fusion as a mode of genetic recombination in are described.


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