Biochemical studies during the development of the chick embryo. VII. Purification and identification of uracil and 5-methylcytosine from chick embryo DNA.

Abstract

It was found that minor components were approximately 1% of the total deoxyribosides in the analysis for base composition of chick embryo DNA. In order to identify the minor components, this work was done. The separation of the minor components from the major components in a DNA hydrolysate was carried out by celite column chromatography and the final purification was accomplished by paper chromatography. For detection of minor components at the deoxyriboside level, the microbial assay method with Lactobacillus leichmannii ATCC 7830 was applied. Thus, two minor deoxyribosides of the DNA were identical with deoxyuridine and 5-methyldeoxycytidine in spectral, electrophoretic and chromatographic properties, respectively. Furthermore, uracil and 5-methylcytosine were purified and identified from perchloric acid hydrolysate of the DNA.