Differences in DNA methylation patterns are detectable during the dimorphic transition of fungi by amplification of restriction polymorphisms (original) (raw)
Abstract
A modification of the amplified fragment length polymorphism technique was developed for the determination of DNA methylation in dimorphic fungi representative of three of the major fungal taxa: Mucor rouxii, a zygomycete; Yarrowia lipolytica, an ascomycete; and Ustilago maydis, a basidiomycete. DNA obtained from the yeast or mycelial stages of the fungi was digested with a mixture of _Eco_RI, and one of the isoschizomers _Msp_I and _Hpa_II, whose ability to cleave at the sequence CpCpGpG is affected by the methylation state. The resulting fragments were ligated to primers and subjected to a double round of amplification by the polymerase chain reaction, radiolabeled in the second round, and separated by polyacrylamide gel electrophoresis. Comparison of patterns revealed differences indicative of fragments whose methylation state did or did not change during the dimorphic transition. These results indicate the usefulness of the method for the study of DNA methylation, demonstrate the universality of DNA methylation in fungi, and confirm that differential DNA methylation occurs during fungal morphogenesis.
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Authors and Affiliations
- Departamento de Ingeniería Genética, Unidad Irapuato, Centro de Investigación y de Estudios Avanzados del IPN, Apartado Postal 629, Irapuato, Gto. 36500, México, , , , ,
G. E. Reyna-López, J. Simpson & J. Ruiz-Herrera
Authors
- G. E. Reyna-López
- J. Simpson
- J. Ruiz-Herrera
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Received: 3 May 1996 / Accepted: 19 September 1996
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Reyna-López, G., Simpson, J. & Ruiz-Herrera, J. Differences in DNA methylation patterns are detectable during the dimorphic transition of fungi by amplification of restriction polymorphisms.Mol Gen Genet 253, 703–710 (1997). https://doi.org/10.1007/s004380050374
- Issue date: February 1997
- DOI: https://doi.org/10.1007/s004380050374