Unexpected substrate specificity of T4 DNA ligase revealed by in vitro selection.
We have used in vitro selection techniques to characterize DNA sequences that are ligated efficiently by T4 DNA ligase. We find that the ensemble of selected sequences ligates about 50 times as efficiently as the random mixture of sequences used as the input for selection. Surprisingly many of the selected sequences failed to produce a match at or close to the ligation junction. None of the 20 selected oligomers that we sequenced produced a match two bases upstream from the ligation junction.
Harada
K
K
Salk Institute for Biological Studies, San Diego, CA 92186-5800.
Orgel
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D001483:Base Sequence; D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D011088:DNA Ligases / Q000378:metabolism*; D008274:Magnesium / Q000494:pharmacology; D008969:Molecular Sequence Data; D013379:Substrate Specificity
D004247:DNA; D011088:DNA Ligases; D008274:Magnesium