Temperature dependence of the joining by T4 DNA ligase of termini produced by type II restriction endonucleases.
The temperature dependence of the T4 DNA ligase-catalyzed joining of plasmid DNA linearized by the action of HaeIII, EcoRI and PstI restriction endonucleases has been investigated by electron microscopy analysis. The extent of joining is maximal at 4 degrees and decreases with increasing temperatures following sigmoid-like curves. The temperature at which 50% of the maximal reaction is still observable increases going from DNA termini without single-stranded overlaps (produced by HaeIII) to termini with four nucleotides overlap, composed only by two A and two T (produced by EcoRI) to termini with four nucleotide overlap, composed by A, T, G and C (produced by PstI).
D011088:DNA Ligases / Q000378:metabolism*; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D005814:Genes, Viral; D007700:Kinetics; D008854:Microscopy, Electron; D010957:Plasmids; D011118:Polynucleotide Ligases / Q000378:metabolism*; D013604:T-Phages / Q000201:enzymology*; D013696:Temperature
D004262:DNA Restriction Enzymes; D011088:DNA Ligases; D011118:Polynucleotide Ligases