title: Development of cloning vectors and transformation methods for Amycolatopsis.
abstract: The genus Amycolatopsis is of industrial importance, as its species are known to produce commercial antibiotics. It belongs to the family Pseudonocardiaceae and has an eventful taxonomic history. Initially strains were identified as Streptomyces, then later as Nocardia. However, based on biochemical, morphological and molecular features, the genus Amycolatopsis, containing seventeen species, was created. The development of molecular genetic techniques for this group has been slow. The scarcity of molecular genetic tools including stable plasmids, antibiotic resistance markers, transposons, reporter genes, cloning vectors, and high efficiency transformation protocols has made progress slow, but efforts in the past decade have led to the development of cloning vectors and transformation methods for these organisms. Some of the cloning vectors have broad host range (pRL series) whereas others have limited host range (pMEA300 and pMEA100). The cloning vector pMEA300 has been completely sequenced, while only the minimal replicon (pA- rep) has been sequenced from pRL plasmids. Direct transformation of mycelia and electroporation are the most widely applicable methods for transforming species of Amycolatopsis. Conjugational transfer from Escherichia coli has been reported only in the species A. japonicum, and gene disruption and replacements using homologous recombination are now possible in some strains.
authors.affiliation: Molecular Biology Laboratory, University of Delhi, Department of Zoology, Delhi 110007, India.
authors.forename: Gauri
authors.initials: G
authors.lastname: Dhingra
authors.forename: Rekha
authors.initials: R
authors.lastname: Kumari
authors.forename: Shashi
authors.initials: S
authors.lastname: Bala
authors.forename: Swati
authors.initials: S
authors.lastname: Majumdar
authors.forename: Shweta
authors.initials: S
authors.lastname: Malhotra
authors.forename: Poonam
authors.initials: P
authors.lastname: Sharma
authors.forename: Sukanya
authors.initials: S
authors.lastname: Lal
authors.forename: John
authors.initials: J
authors.lastname: Cullum
authors.forename: Rup
authors.initials: R
authors.lastname: Lal
mesh_terms: D000192:Actinomycetales / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D018014:Gene Transfer Techniques; D007218:Industrial Microbiology / Q000379:methods*; D010957:Plasmids / Q000235:genetics; D014169:Transformation, Bacterial