USE OF RANDOM SYSTEMATIC MUTAGENESIS TO GENERATE VIABLE HUMAN RHINOVIRUS 14 CHIMERAS DISPLAYING HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 V3 LOOPSEQUENCES

Random systematic mutagenesis was used to generate a library of human rhinovirus 14 chimeras that each display a segment from the V3 loop of human immunodeficiency virus type 1. The sequence XXIGPGRAXX, where X could be any of the 20 amino acids, was inserted at the neutralizing immunogenic site II of human rhinovirus 14 between VP2 residues 159 an d 160. Twenty-five unique chimeric viruses were isolated, and the iden tity of their randomized residues was determined. A nonrandom amino ac id distribution that may reflect structural requirements for viability was observed at the randomized positions. Fifteen of 25 chimeras were neutralized by one or more of a panel of four anti-human immunodefici ency virus type 1 V3 loop antibody preparations, indicating that antig enicity had been successfully transplanted. Libraries of chimeric viru ses produced by using the techniques described may be a source of vacc ines and other immunotherapeutic reagents. The random systematic mutag enesis methodology described should be generally useful for the rapid transplantation of foreign sequences into viral coat and other protein s to produce libraries containing members with the desired properties.