PMID: 6652091Nov 17, 1983Paper

Ligation of highly modified bacteriophage DNA

Biochimica Et Biophysica Acta
X M CaoM Ehrlich

Abstract

After digestion by TaqI or nicking by DNAase I, five highly modified bacteriophage DNAs were tested as substrates for T4 DNA ligase. The DNAs used were from phages T4, XP12, PBS1, SP82, and SP15, which contain as a major base either glucosylated 5-hydroxymethylcytosine, 5-methylcytosine, uracil, 5-hydroxymethyluracil, or phosphoglucuronated, glucosylated 5-(4',5'-dihydroxypentyl)uracil, respectively. The relative ability of cohesive-ended TaqI fragments of these DNAs and of normal, lambda DNA to be ligated was as follows: lambda DNA = XP12 DNA greater than SP82 DNA approximately equal to nonglucosylated T4 DNA greater than T4 DNA = PBSI1 DNA much greater than SP15 DNA. Taq I-T4 DNA fragments were also inefficiently ligated by Escherichia coli DNA ligase. However, annealing-independent ligation of DNAase I-nicked T4, PBS1, and lambda DNAs was equally efficient. We conclude that the poor ligation of Taq I fragments of T4 and PBS1 DNAs was due to the hydroxymethylation (and glucosylation) of cytosine residues at T4's cohesive ends and the substitution of uracil residues for thymine residues adjacent to PBS1's cohesive ends destabilizing the annealing of the restriction fragments. Only SP15 DNA with its negatively charged, modified...Continue Reading

References

Jan 4, 1966·Biochemical and Biophysical Research Communications·T L UlbrichtI Takahashi
Aug 1, 1970·European Journal of Biochemistry·N TruffautM O Soulie
Mar 11, 1982·Nucleic Acids Research·L H HuangM Ehrlich
Jan 1, 1980·Annual Review of Microbiology·R A Warren

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Citations

Aug 31, 2018·Molecular Microbiology·Yifeng YuanValérie de Crécy-Lagard
Aug 23, 2017·Scientific Reports·Rebecca TsaiShuang-Yong Xu

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