Abstract
Seventeen species of 11 rare actinomycete genera were treated with lysozyme and achromopeptidase to produce protoplasts. Fourteen of them could be protoplasted. Abundant protoplasts were obtained from Actinokineospora globicatena, Actinosynnema mirum and Amycolatopsis azurea, while relatively small amount of protoplasts were obtained from two Actinomadura species and Couchioplanes caeruleus subsp. azureus. Nine of 14 protoplasted species could regenerate on R1M and R2YE media with the range of regeneration frequency. The protoplasts of Amycolatopsis azurea regenerated on R1M and R2YE at the highest frequency (80%). Actinomadura atramentaria, A. echinospora, Catellatospora ferruginea and two Couchioplanes species could not regenerate on any tested media. A. azurea could be transformed with a Streptomyces vector, pIJ702, at the frequency of 1.6 x 106 transformants per μg DNA. The other Streptomyces plasmid pRES18 was also able to be maintained in this strain.
