By replacing radiation with fluorescent system in the technique of methylation sensitive amplified polymorphism (MSAP) and optimizing reaction conditions, a modified technique to detect DNA methylation called F-MSAP (fluorescent labeled methylation sensitive amplified polymorphism) was developed. In the present study, cytosine methylation patterns of genomic DNA were investigated in two inbred chickens and their F1 hybrids. Three types of methylation patterns were observed in each individual, namely fully methylated, hemi-methylated or not methylated types. The average incidence of methylation was approximately 40%. The percentage that the F1 hybrid individual inherits the methylation for any given sites from either/both parent amounted to 95%, while the percentage of altered methylation patterns in F1 individual was only 5%, including 14 increased and 12 decreased methylation types, demonstrating that F-MSAP was highly efficient for large-scale detection of cytosine methylation in chicken genome. Our technique can be further extended to other animals or plants with complex genome and rich in methylation polymorphism.
This is a preview of subscription content, log in to check access.
Buy single article
Instant unlimited access to the full article PDF.
Price includes VAT for USA
Courtier, B., Heard, E., Avner, P., Xce haplotypes show modified methylation in a region of the active X chromosome lying 3′ to Xst, Proc. Natl. Acad. Sci. USA, 1995, 92(n8): 3531–3535.
Thorvaldsen, J. L., Duran, K. L., Bartolomei, M. S., Deletion of the H19 differentially methylated domain results in loss of imprinted expression of H19 and Igf2, Genes Dev., 1998, 12(n23): 3693–3702.
Momparler, R. L., Bovenzi, V., DNA methylation and cancer, J. Cell Physiol., 2000, 183(n2): 145–154.
Bird, A. P., Taggart, M. H., Variable patterns of total DNA and rDNA methylation in animals, Nucleic Acids Res., 1980, 8(n7): 1485–1497.
Herman, J. G., Graff, J. R., Nelkin, B. D. et al., Methyla-tion-specific PCR: A novel PCR assay for methylation status of CpG islands, Proc. Natl. Acad. Sci. USA, 1996, 93(n18): 9821–9826.
Adorjan, P., Distler, J., Lipscher, E. et al., Tumor class prediction and discovery by microarray-based DNA methylation analysis, Nu-cleic Acids Res., 2002, 30(n5): e21.
Xiong, L. Z., Xu, C. G., Zhang, Q. et al., Patterns of cytosine me-thylation in an elite rice hybrid and its parental lines, detected by a methylation-sensitive amplification polymorphism technique, Mol. Gen. Genet., 1999, 261(n3): 439–446.
Labra, M., Grassi, F., Imazio, S. et al., Genetic and DNA-methylation changes induced by potassium dichromate in Brassica napus L., Chemosphere, 2004, 54(n8): 1049–1058.
Aina, R., Sgobati, S., Santagotino, A. et al., Specific hypomethyla-tion of DNA is induced by heavy metals in white clover and indus-trial hemp, Physiol. Plant, 2004, 121(n3): 472–480.
Li, X. Q., Xu, M. L., Korban, S. S., DNA methylation profiles dif-fer between field-and in vitro-grown leaves of apple, Plant Physiol., 2002, 159(n11): 1229–1234.
de Montera, B., Boulanger, L., Taourit, S. et al., Genetic identity of clones and methods to explore DNA, Cloning Stem Cells, 2004, 6(n2): 133–139.
Shaked, H., Kashush, K., Ozkan, H. et al., Sequence elimination and cytosine methylation are rapid and reproducible responses of the genome to wide hybridization and allopolyploidy in wheat, Plant Cell, 2001, 13(n8): 1749–1759.
Madlung, A., Masuell, R. W., Watson, B. et al., Remodeling of DNA methylation and transcriptional changes in synthetic Arabi-dopsis allotetraploids, Plant Physiol., 2002, 129(n2): 733–746.
Xu, M. L., Li, X. Q., Korban, S. S., AFLP-based detection of DNA methylation, Plant Mol. Biol. Rep., 2000, 18(n4): 361–368.
Vos, P., Hogers, R., Bleeker, M. et al., AFLP: A new technique for DNA fingerprinting, Nucleic Acids Res., 1995, 23(n21): 4407–4414.
Huang, J. C., Sun, M., A modified AFLP with fluorescence-labelled primers and automated DNA sequencer detection for efficient fin-gerprinting analysis in plants, Biotechnol. Tech., 1999, 13(n4): 277–278.
Zhao, S., Mitchell, S. E., Meng, J. et al., Genomic typing of Es-cherichia coli O157: H7 by semi-automated fluorescent AFLP analysis, Microbes Infect., 2000, 2(n2): 107–113.
Terefework, Z., Kaijalainen, S., Lindstrom, K., AFLP fingerprint-ing as a tool to study the genetic diversity of Rhizobium galegae isolated from Galega orientalis and Galega officinalis, J. Biotech-nol., 2001, 91(n2–3): 169–180.
McClelland, M., Nelson, M., Raschke, E., Effect of site-specific modification on restriction endonucleases and DNA modification methyltransferases, Nucleic Acids Res., 1994, 22(n17): 3640–3659.
About this article
Cite this article
Xu, Q., Sun, D. & Zhang, Y. F-MSAP: A practical system to detect methylation in chicken genome. Chin.Sci.Bull. 50, 2039–2044 (2005). https://doi.org/10.1007/BF03322798
- methylation sensitive amplified polymorphism
- fluorescent labeled
- DNA methylation