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Molecular characterization of a gene from Saccharopolyspora erythraea (Streptomyces erythraeus) which is involved in erythromycin biosynthesis
Author(s) -
Dhillon N.,
Hale R. S.,
Cortes J.,
Leadlay P. F.
Publication year - 1989
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1989.tb00123.x
Subject(s) - biology , gene cluster , streptomyces , gene , shuttle vector , genetics , plasmid , nucleic acid sequence , mutant , streptomycetaceae , promoter , actinomycetales , recombinant dna , gene expression , bacteria , vector (molecular biology)
Summary A 7.3 kbp DNA fragment, encompassing the erythromycin (Em) resistance gene ( ermE ) and a portion of the gene cluster encoding the biosynthetic genes for erythromycin biosynthesis in Saccharopolyspora erythraea (formerly Streptomyces erythraeus ) has been cloned in Streptomyces lividans using the plasmid vector pIJ702, and its nucleotide sequence has been determined using a modified dideoxy chain‐termination procedure. In particular, we have examined the region immediately 5′ of the resistance determinant, where the tandem promoters for ermE overlap the promoters for a divergently transcribed coding sequence (ORF). Disruption of this ORF using an integrational pIJ702‐based plasmid vector gave mutants which were specifically blocked in erythromycin biosynthesis, and which accumulated 3‐O‐α‐L‐mycarosylerythronolide B: this behaviour is identical to that of previously described eryC 1 mutants. The eryC 1‐gene product, a protein of subunit M r 39200, is therefore involved either as a structural or as a regulatory gene in the formation of the deoxyamino‐sugar desosamine or in its attachment to the macro‐lide ring.