Development of cloning vectors and transformation methods for<Emphasis Type="Italic"> Amycolatopsis</Emphasis> |
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Authors: | Gauri?Dhingra Rekha?Kumari Shashi?Bala Swati?Majumdar Shweta?Malhotra Poonam?Sharma Sukanya?Lal John?Cullum Email author" target="_blank">Rup?LalEmail author |
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Institution: | (1) Molecular Biology Laboratory, University of Delhi, Department of Zoology, Delhi 110007, India,;(2) LB Genetik, University of Kaiserslautern, Fachbereich Biologie, Postfach 3049, 67663, Kaiserslautern, Germany, |
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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.
Electronic Publication |
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Keywords: | Amycolatopsis Plasmids Cloning vectors Marker genes Transformation Replicon |
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