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Formation of a minichromosome in Cryptococcus neoformans as a result of electroporative transformation

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Abstract

A minichromosome of approximately 270 kilobases was generated following complementation of a ura5 mutant strain of C. neoformans with the plasmid pURA5g2. This is the first report of the in-vivo generation of a minichromosome by the method of electroporative transformation. The minichromosome occurred at a relatively high (>20%) frequency in transformants that were stable for uracil protoprophy. The minichromosome was maintained in linear form as a large extrachromosomal element of the normal karyotype. Gel-purified DNA from the minichromosome readily transformed the ura5 mutant of C. neoformans. Southern-blot analysis of the minichromosome revealed the presence of multiple copies of the URA5 gene and ribosomal DNA sequences in addition to containing telomere-like sequence repeats. The minichromosome was transmitted through mitosis and meiosis with extremely-high fidelity.

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Authors and Affiliations

  1. Clinical Mycology Section, Laboratory-of Clinical Investigation, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 20892, Bethesda, MD, USA

    Ashok Varma & K. J. Kwon-Chung

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  1. Ashok Varma
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  2. K. J. Kwon-Chung
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Communicated by H. Bertrand

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Varma, A., Kwon-Chung, K.J. Formation of a minichromosome in Cryptococcus neoformans as a result of electroporative transformation. Curr Genet 26, 54–61 (1994). https://doi.org/10.1007/BF00326305

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  • DOI: https://doi.org/10.1007/BF00326305

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