Abstract
The carboxy-terminal domain of polymerase gene of Rous sarcoma virus was cloned into an expression vector under the control oflac regulatory elements, resulting in the plasmid pMF1413. Upon isopropyl-β-D-thiogalactopyranoside induction, viral integration (IN) protein was expressed in large quantity inEscherichia coli. The expressed recombinant protein was prepurified by successive washing of the bacterial pellet with 0.1 M NaCl and detergents. Further purification was performed in high yield by standard chromatography methods. The purified enzyme revealed selective DNA cleaving activity on supercoiled plasmid with the LTR-LTR junction fragment. The reaction was metal ion dependent, with a preference for Mn2+ over Mg2+, and showed substrate specificity at 1 mM MnCl2.
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Marczinovits, I., Molnár, J., Sóki, J. et al. Overexpression and purification of enzymatically active recombinant integrase protein of rous sarcoma virus. Virus Genes 6, 301–306 (1992). https://doi.org/10.1007/BF01702568
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DOI: https://doi.org/10.1007/BF01702568