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Nucleic Acids Research, 1981, Vol. 9, No. 1 85-93
© 1981

ENZYMOLOGY

Temperature dependence of the joining by T4 DNA ligase of termini produced by type II restriction endonucleases

Luca Ferretti and Vittorio Sgaramella

Istituto di Genetica Biochimica ed Evoluzionistica del C.N.R. Via S.Epifanio, 14, 27100 Pavia, Italy

Received October 8, 1980. 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° 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)


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