Determination of 5-methylcytosine from plant DNA by high-performance liquid chromatography

Journal/Book: Biochim-Biophys-Acta 654 (1), 52-56. 1981;

Abstract: The relative amounts of the five nucleosides (deoxycytidine,5-methyldeoxycytidine, deoxyadenosine, deoxyguanosine and thymidine) inthe DNA of nine plant species, one plant satellite DNA, and one animalspecies were determined by high performance liquid chromatography. Themethod allows the clean separation of the nucleosides from 10 microgramsamples with 15 min. The following values for the proportion ofmethylated cytosines among all cytosines were obtained: Lobulariamaritima 18.5%, Nicotiana tabacum 32.6%, Pisum sativum 23.2%, Rhinanthusminor 29.2%, Sinapsis alba 12.2%, Vicia faba 30.5%, Viscum album 23.2%,Cymbidium pumilum 18.8%, Cymbidium pumilum AT-rich satellite DNA 15.8%,Triticum aestivum 22.4%. DNA of an animal, the gerbil, Merionesunguiculatus, had a methylation percentage of 3.1%. An estimate of theGC content based on the buoyant density of DNA tends to be lower thanthe actual value, an estimate based on the melting temperature tends tobe higher. This supports the finding by other authors that DNAmethylation decreases the buoyant density and may increase the meltingtemperature at high m5C concentration. Author.

Keyword(s): CHROMATOGRAPHY-HIGH-PRESSURE-LIQUID/MT (methods)

Zurück | Weiter