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Summary: Twenty-seven mutants defective in the biosynthesis of biotin were generated by N-methyl-N -nitro-N-nitrosoguanidine treatment of the obligate methylotrophic bacterium ‘Methylobacillus flagellatum’. The metabolic lesion in some of these mutants was determined in cross-feeding experiments using known Escherichia coli bio mutants. R-prime plasmids carrying various fragments of the ‘M. flagellatum’ chromosome were used for complementation of bio mutations in ‘M. flagellatum’ and E. coli. By complementation analysis and cross-feeding experiments, ‘M. flagellatum’ Bio- mutants were classified into five groups: bioA, bioB, bioD, bioH, and a group with unidentified bio mutations (bioF or/and bioC). Using R-prime complementation mapping and also Hfr-like mapping, most of the bio genes of ‘M. flagellatum’ identified in this study were localized on the ‘M. flagellatum’ chromosome.
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