Development of a Fluorometric In Vitro Assay for HIV

The function of retroviral integrase (IN) is the integration of viral DNA into host cell DNA, an esretroviral replication. IN protein recognizes viral cognate at sites, removes two nucleotides from tviral DNA (3' processing) and joins the 3'-processed cognate at sites to the newly created 5' ends otransfer). This research will develop a fluorometric assay utilizing fluorescence resonance energy tprimarily the result of dipole-dipole interactions between the donor and the acceptor. Hence, when afluorophores are introduced to a DNA substrate in close proximity, the donor fluorescence will be quThe process will be fully reversed if fluorophores are physically separated by enzyme-mediated DNA cquenched fluorescence will be recovered and the extent of this change upon cleavage of fluorescent Dreflect the extent of the cleavage reaction of DNA. This assay is non radioactive, sensitive, rapid,possible to obtain precise kinetic information essential for mechanistic studies and screening of en