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PMID: 2987859 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

alpha-Putrescinylthymine and the sensitivity of bacteriophage phi W-14 DNA to restriction endonucleases.

Nucleic acids research ·Vol. 13 ·No. 7 ·1985-04-11 ·Pages 2559-68

Miller PB, Wakarchuk WW, Warren RA

Abstract

The modified base alpha-putrescinylthymine (putT) in phi W-14 DNA blocks cleavage of the DNA by 17 of 32 Type II restriction endonucleases. The enzymes cleaving the DNA do so to widely varying extents. The frequencies of cleavage of three altered forms of the DNA show that putT blocks recognition sites either when it occurs within the site or when it is in a sequence flanking the site. The blocking is dependent on both charge and steric factors. The charge effects can be greater than the steric effects for some of the enzymes tested. All the enzymes cleaving phi W-14 DNA release discrete fragments, showing that the distribution of putT is ordered. The cleavage frequencies for different enzymes suggest that the sequence CAputTG occurs frequently in the DNA. Only TaqI of the enzymes tested appeared not to be blocked by putT, but it was slowed down. TaqI generated fragments are joinable by T4 DNA ligase.

MeSH Terms
Bacteriophages/genetics Base Sequence Binding Sites DNA Restriction Enzymes/metabolism DNA, Viral/analysis Deoxyribonucleases, Type II Site-Specific Electrophoresis, Agar Gel Thymine/analogs & derivatives,pharmacology
Chemicals
DNA, Viral alpha-putrescinylthymine DNA Restriction Enzymes Deoxyribonucleases, Type II Site-Specific TCGA-specific type II deoxyribonucleases Thymine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Miller P B
Wakarchuk W W
Warren R A
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35 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/13.7.2559
Published
1985-04-11
Pages
2559-68
Language
English
Region
England
NLM ID
0411011
PMCID
PMC341175
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/13.7.2559 PMC Full Text View journal details
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