Molecular and General Genetics MGG

, Volume 253, Issue 6, pp 703–710

Differences in DNA methylation patterns are detectable during the dimorphic transition of fungi by amplification of restriction polymorphisms

  • G. E. Reyna-López
  • J. Simpson
  • J. Ruiz-Herrera
ORIGINAL PAPER

DOI: 10.1007/s004380050374

Cite this article as:
Reyna-López, G., Simpson, J. & Ruiz-Herrera, J. Mol Gen Genet (1997) 253: 703. doi:10.1007/s004380050374

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 EcoRI, and one of the isoschizomers MspI and HpaII, 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.

Keywords Amplified fragment length polymorphism  Fungal dimorphism   DNA methylation 

Copyright information

© Springer-Verlag Berlin Heidelberg 1997

Authors and Affiliations

  • G. E. Reyna-López
    • 1
  • J. Simpson
    • 1
  • J. Ruiz-Herrera
    • 1
  1. 1.Departamento de Ingeniería Genética, Unidad Irapuato, Centro de Investigación y de Estudios Avanzados del IPN, Apartado Postal 629, Irapuato, Gto. 36500, México
  2. 2.Departamento de Genética y Biología Molecular, Centro de Investigacíon y de Estudios Avanzados del IPN, México, D. F., México
  3. 3.Facultad de Química, Universidad Autónoma de Guanajuato, México

Personalised recommendations