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Characterization of human DNA ligase I expressed in a baculovirus-insect cell system
A Gallina1, F Rossi, G Milanesi
1Istituto di Genetica Biochimica ed Evoluzionistica, CNR, Pavia, Italy.
The Biochemical Journal
|December 1, 1995
Abstract:
The baculovirus expression system was used to overexpress human DNA ligase I (hLig I). Approx. 2 mg of recombinant hLig I was produced per 10(8) Spodoptera frugiperda Sf9 insect cells infected with the recombinant baculovirus. The optimum time point for the production of biologically active recombinant hLig I was 48 h post-infection. Lig I activity was demonstrated by auto-adenylating, polynucleotide joining and DNA relaxation assays. The baculovirus system has the advantage over previously described methods for producing hLig I of generating large amounts of a full-length active protein.