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Development of cloning vectors and transformation methods for Amycolatopsis

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Journal of Industrial Microbiology and Biotechnology

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.

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Acknowledgements

The project was supported in part by grants from the Department of Science and Technology, Government of India. R. Kumari, Swati Majumdar, Shweta Malhotra and Poonam Sharma gratefully acknowledge the Council of Scientific and Industrial Research (CSIR), and the University Grants Commission (UGC), respectively, for providing doctoral fellowships. Rup Lal and John Cullum thank the Department of Science and Technology (Govt. of India) and the Deutsche Akademische Austauschdienst (DAAD), Germany, for support under the project-based personnel exchange programme.

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Authors and Affiliations

  1. Molecular Biology Laboratory, University of Delhi, Department of Zoology, Delhi 110007, India

    Gauri Dhingra, Rekha Kumari, Shashi Bala, Swati Majumdar, Shweta Malhotra, Poonam Sharma, Sukanya Lal & Rup Lal

  2. LB Genetik, University of Kaiserslautern, Fachbereich Biologie, Postfach 3049, 67663, Kaiserslautern, Germany

    John Cullum

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  1. Gauri Dhingra
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Correspondence to Rup Lal.

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Dhingra, G., Kumari, R., Bala, S. et al. Development of cloning vectors and transformation methods for Amycolatopsis . J IND MICROBIOL BIOTECHNOL 30, 195–204 (2003). https://doi.org/10.1007/s10295-003-0040-6

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  • DOI: https://doi.org/10.1007/s10295-003-0040-6

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