Transformation of Arthrobacter and studies on the transcription of the Arthrobacter ermA gene in Streptomyces lividans and Escherichia coli

Abstract

We report the development of a plasmid-mediated transformation system for Arthrobacter sp. NRRLB3381, using the Streptomyces cloning vector pIJ702. Our procedure gives a transformation frequency of 10(3)/micrograms of plasmid DNA. In addition we have explored the expression of the Arthrobacter ermA gene in Streptomyces lividans and Escherichia coli, and shown that the ermA promoter is recognized in S. lividans not E. coli. The relationship between Arthrobacter, Streptomyces and E. coli promoters is discussed.

Description of the Erythromycin-Producing Bacterium Arthrobacter sp. Strain NRRL B-3381 as Aeromicrobium erythreum gen. nov., sp. nov.

“…strain NRRL B-3381T differs from most macrolide-producing bacteria in that it is nonsporulating and produces erythromycin throughout growth, and the only form of erythromycin produced is erythromycin A. These unique characteristics have prompted studies of gene structure and regulation in strain NRRL B-3381T (14,22,23).…”

The erythromycin biosynthetic gene cluster of Aeromicrobium erythreum

“…The unicellular actinomycete, Aeromicrobium erythreum, however also produces erythromycin [13]. Previous studies with A. erythreum by Roberts et al [31,32] focused on the development of a transformation system and characterization of the erythromycin resistance gene ermR (formerly ermA). A. erythreum was assigned to the genus Aeromicrobium by Miller et al [23], replacing its former classification in the genus Arthrobacter .…”

Plasmids for Nicotine-Dependent and -Independent Gene Expression in Arthrobacter nicotinovorans and Other Arthrobacter Species

“…Most of the Arthrobacter genes characterized so far have been cloned and expressed in Escherichia coli (10,(15)(16)(17) and other bacterial hosts (16,18,19), or alternatively, the proteins have been purified directly from crude lysates of Arthrobacter. Heterologous expression, however, can lead to inactive proteins due to misfolding, a lack of appropriate enzyme cofactors and proper insertion of these cofactors into apoenzymes, or posttranslational modifications.…”