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Transformation of Mucor circinelloides with Autoreplicative Vectors Containing Homologous and Heterologous ARS Elements and the Dominant Cbxr Carboxine-Resistance Gene

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Abstract

Mucor circinelloides transformants prototrophic to leucine and resistant to carboxine (Leu+ Cbxr) have been obtained by treatment of protoplasts with plasmid constructs containing homologous leuA gene and adjacent autonomously replicating sequences (ARS) element combined with the Cbxr(carboxine-resistance) gene of Ustilago maydis and ARS sequences from this basidiomycete (plasmid pGG37) or from the 2 μ plasmid of Saccharomyces cerevisiae (plasmid pGG43). The presence in the same plasmid molecule of the M. circinelloides leuA gene and adjacent ARS element together with heterologous ARS elements produced an increase in the transformation frequency of about 65–120%. The presence of autoreplicating plasmid molecules in the transformants was demonstrated by mitotic stability experiments, by Southern analysis, and by the rescue of plasmids from transformed bacterial cells.

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Acknowledgments

This work was supported by Consejo Nacional de Ciencia y Tecnología (CONACyT) and Universidad de Guanajuato, Mexico. R.O.A. received a fellowship from CONACyT, Mexico.

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Authors and Affiliations

  1. Instituto de Investigación en Biología Experimental, Facultad de Química, Universidad de Guanajuato, Noria Alta s/n, Apartado Postal 187, Guanajuato, Gto., 36000, México

    R. Ortiz-Alvarado, G. A. Gonzalez-Hernandez, J. C. Torres-Guzman & J. F. Gutierrez-Corona

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  1. R. Ortiz-Alvarado
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  2. G. A. Gonzalez-Hernandez
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  3. J. C. Torres-Guzman
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  4. J. F. Gutierrez-Corona
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Correspondence to J. F. Gutierrez-Corona.

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Ortiz-Alvarado, R., Gonzalez-Hernandez, G.A., Torres-Guzman, J.C. et al. Transformation of Mucor circinelloides with Autoreplicative Vectors Containing Homologous and Heterologous ARS Elements and the Dominant Cbxr Carboxine-Resistance Gene. Curr Microbiol 52, 178–181 (2006). https://doi.org/10.1007/s00284-005-0088-9

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  • DOI: https://doi.org/10.1007/s00284-005-0088-9

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