A physical map of the chromosome of the erythromycin-producing actinomycete NRRL 2338 has been constructed using the restriction enzymes and . The map was constructed by a variety of methods including linking clone analysis, cross-hybridizations using labelled macrorestriction fragments, gene probing, two-dimensional PFGE and restriction enzyme site generation. Analysis of the individual macrorestriction patterns of the 17 -, 6 - and 22 -digested fragments indicated a chromosome size of about 8 Mb. Linking clones for five contiguous fragments were obtained, covering 32% of the chromosome. The linkage of an additional eight fragments was aided by the finding that the rRNA operons of contain an site within the 16S gene. Generation of strains that contain additional sites within selected fragments, followed by PFGE analysis and Southern hybridization to determine specific linkages, facilitated the completion of the map. The entire map was constructed by gene probing and cross-hybridizations. PFGE analysis of agarose-embedded DNA prepared in either the presence or absence of proteinase K suggested that the NRRL 2338 chromosome is linear. A total of 15 genes or gene clusters were mapped to specific and fragments, including the erythromycin-biosynthetic gene cluster and the rRNA operons.


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