A multicopy vector system for genetic studies in Mucor circinelloides and other zygomycetes View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2004-06

AUTHORS

K. F. Appel, A. M. Wolff, J. Arnau

ABSTRACT

Transformation of Mucor circinelloides is routinely achieved by using a plasmid containing the wild-type leuA gene to complement the leucine requirement of an auxotrophic host strain. As is the case for other zygomycetes, the transforming DNA is usually not integrated into the genome of M. circinelloides, but is maintained as an autonomously replicating plasmid. However, even under selective conditions, the plasmid is segregationally unstable, resulting in a rather low number of cells carrying the plasmid. We report here on a new transformation vector based on a dominant selection marker conferring resistance to geneticin, which allows for plasmid maintenance in high copy numbers. The vector was also used to transform Mucor rouxii and Rhizomucor pusillus, and should therefore be a valuable tool for gene expression studies in zygomycetes. The functionality and regulatory properties of the promoter of the M. circinelloides gpd1 gene (which codes for glyceraldehyde-3P-dehydrogenase) were demonstrated in R. pusillus using geneticin selection. In this work, we have also determined the molecular basis of the Leu(-) phenotype of the M. circinelloides host strain R7B. The leucine requirement is due to a single point mutation in the leuA gene that results in the replacement of a glutamic acid by a lysine residue. More... »

PAGES

595-602

References to SciGraph publications

Journal

TITLE

Molecular Genetics and Genomics

ISSUE

5

VOLUME

271

Author Affiliations

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/s00438-004-1008-6

    DOI

    http://dx.doi.org/10.1007/s00438-004-1008-6

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1048729186

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/15088140


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