Dynemicin is a novel anthraquinone-fused member of the 10-membered enediyne antitumor antibiotic family. The development of a genetic system for the dynemicin producer Micromonospora chersina confirmed, for the first time, the requirement of the putative enediyne core biosynthetic genes (dynE8, U14 and U15) and a tailoring oxidase gene (orf23) for dynemicin production. Cloning and sequence analysis of a 76 kb of genomic sequence region containing dynE8 revealed a variety of genes conserved among known enediyne loci. Surprisingly, this fragment and flanking chromosomal DNA lacked any obvious genes encoding for the biosynthesis of the anthraquinone, suggesting that the location of genes encoding for the biosynthesis of the dynemicin enediyne core and the dynemicin anthraquinone are chromosomally distinct. The demonstrated trace production of a shunt product from mutant strain QGD23 (Deltaorf23) also sets the stage for subsequent studies to delineate the key steps in enediyne core biosynthesis and tailoring.