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October 6, 2022 12:00
Sala d’Actes IQAC
The reseracher Dr. Martin Willemoës, from the Linderstrøm-Lang Center of the University of Copenhagen, will give an IQAC Seminar, invited by Dr. Ramon Crehuet, from the Theoretical and Computational Chemistry group.
Date: Thursday 6th October at 12:00h.
Title: Characterizing HIV protease and variants utilizing an Escherichia coli genetic system.
Abstract:
A genetic selection system for activity of HIV protease is described that is based on a synthetic substrate constructed as a modified AraC regulatory protein that when cleaved stimulate L-arabinose metabolism in an Escherichia coli araC strain. Growth stimulation on selective plates was shown to depend on active HIV protease and the scissile bond in the substrate. In addition, the growth of cells correlated well with the established cleavage efficiency of the sites in the viral polyprotein, Gag, when these sites were individually introduced into the synthetic substate of the selection system. Plasmids encoding protease variants selected based on stimulation of cell growth in the presence of saquinavir or cleavage of a site not cleaved by wild-type protease, were indistinguishable with respect to both phenotypes. Also, both groups of selected plasmids encoded side chain substitutions known from clinical isolates or displayed different side chain substitutions but at identical positions.