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SUMMARY: A chromosomal linkage map of ten markers was established for Streptomyces ambofaciens by the four-factor cross method and allele-gradient analysis. Mutants were obtained by nitrous acid treatment as well as by UV mutagenesis. The fertility of crosses was enhanced over 100-fold by pSAM2, a plasmid present in some strains of S. ambofaciens, and over 1000-fold by the conjugative plasmid pIJ303.