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PMID: 6255419 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The 5'-cytosine in CCGG1 is methylated in two eukaryotic DNAs and Msp I is sensitive to methylation at this site.

Nucleic acids research ·Vol. 8 ·No. 17 ·1980-09-11 ·Pages 3829-40

Sneider TW

Abstract

Novikoff rat hepatoma and bovine liver DNAs were digested with Msp I or Hpa II. Restriction fragments were end-labeled using [alpha-32P]-dCTP and the Klenow fragment of E. coli DNA polymerase I and then digested to 2'-deoxyribonucleoside-3'-monophosphates using micrococcal nuclease and spleen phosphodiesterase. Mononucleotides were separated by two-dimensional thin layer chromatography, localized by radioautography, and the [32P]-label quantitated by scintillation spectrometry. This method, based on known specificities of Msp I and Hpa II, shows that CCGG, CMGG, and MCGG (M refers to 5-methylcytosine) occur at frequencies of 89.6%, 1.4%, and 9.0%, respectively, in the rat DNA and at 41.6%, 48.3%, and 10.0%, respectively, in the bovine DNA. [32P] recovery in 3'-5-MedCMP from end-labeled Msp I digests was negligible compared to recovery from Hpa II digests. Hence, Msp I is sensitive to methylation at the 5' cytosine in the sequence CCGG.

MeSH Terms
5-Methylcytosine Animals Base Sequence Cattle Cytosine/analogs & derivatives,analysis DNA DNA Restriction Enzymes DNA, Neoplasm Deoxyribonucleases Liver/analysis Liver Neoplasms, Experimental/analysis Oligodeoxyribonucleotides/analysis Oligonucleotides/analysis Rats
Chemicals
DNA, Neoplasm Oligodeoxyribonucleotides Oligonucleotides 5-Methylcytosine Cytosine DNA Deoxyribonucleases DNA Restriction Enzymes
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Sneider T W
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22 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/8.17.3829
Published
1980-09-11
Pages
3829-40
Language
English
Region
England
NLM ID
0411011
PMCID
PMC324197
Subset
IM
Grants
NCI NIH HHS · CA-18719 · United States
External Links
PubMed source DOI: 10.1093/nar/8.17.3829 PMC Full Text View journal details
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