Biochemical and cultural analysis of mycobacterial recombinants obtained by spheroplast fusion.
Spheroplast fusion using polyethylene glycol 6000 resulted in mycobacterial recombinants with a frequency of about 10(-3). The parental strains were two carotenoid pigment mutants of Mycobacterium aurum which were obtained by chemical mutagenesis of the wild-type strain with N-methyl-N'-nitro-N-nitrosoguanidine. Isolation of recombinant clones was based on their pigmentation on Löwenstein-Jensen medium, and genetically stable recombinants were analysed for their carotenoid pigments and their alpha- and beta-mycolic acids. They were then submitted to 14 biochemical tests as well as determination of their susceptibility to 13 drugs and their ability to grow at 30 and 42 degrees C. Recombinants were found to be positive for nitrate reductase (100%), gamma-glutamyl transferase (94%), urease (53%) and arylsulphatase (21%). Out of 34 recombinants tested, 5 grew both at 30 degrees and 42 degrees C, whereas 1 grew only at 37 degrees C. The results of drug susceptibility testing were inconclusive.
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001192:Arylsulfatases / Q000032:analysis; D001669:Biochemical Phenomena; D001671:Biochemistry; D002338:Carotenoids / Q000032:analysis; D003470:Culture Media; D004352:Drug Resistance, Microbial; D009154:Mutation; D009161:Mycobacterium / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D009171:Mycolic Acids / Q000032:analysis; D050901:Nitrate Reductase; D009565:Nitrate Reductases / Q000032:analysis; D011995:Recombination, Genetic; D013104:Spheroplasts; D013696:Temperature; D014510:Urease / Q000032:analysis; D005723:gamma-Glutamyltransferase / Q000032:analysis
D000900:Anti-Bacterial Agents; D003470:Culture Media; D009171:Mycolic Acids; D002338:Carotenoids; D009565:Nitrate Reductases; D050901:Nitrate Reductase; D005723:gamma-Glutamyltransferase; D001192:Arylsulfatases; D014510:Urease