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J. Biol. Chem., Vol. 255, Issue 16, 7544-7547, Aug, 1980

Enzymatic analysis of 5-methylcytosine content in eukaryotic DNA. Study of intracellular Simian Virus 40 DNA

JP Ford, M Coca-Prados and MT Hsu

An enzymatic method is described for the analysis of the content of 5- methylcytosine in the DNA of eukaryotic cells. The conventional acid hydrolysis procedure was found to result in variable deamination of 5- methylcytosine into thymine, making the exact quantitation difficult. We applied the enzymatic method for the analysis of a simple eukaryotic viral genome. Viral and cell DNA in Simian Virus 40-infected monkey kidney cells was labeled with [methyl-3H]methionine in vivo and their 5- methylcytosine content was analyzed. The content of 5-methylcytosine per thymine in the total intracellular viral DNA was found to be less than 10(-4) of the host DNA. The implication of this result with regard to eukaryotic gene expression is discussed.
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				M Ehrlich and R. Wang 5-Methylcytosine in eukaryotic DNA Science, June 19, 1981; 212(4501): 1350 - 1357. [Abstract] [PDF]