EFFICIENT RECOVERY AND REGENERATION OF INTEGRATED RETROVIRUSES

We report a rapid and efficient PCR-based rescue procedure for integra ted recombinant. retroviruses. Full-length proviral DNA is amplified b y long-range PCR using a pair of primers derived from the long termina l repeats (LTR), and virus is regenerated by transfecting retrovirus-p ackaging cells with the PCR-derived provirus. The viral yield from the PCR product is similar to that from the retroviral plasmid vector, an d the representation of different inserts is accurately maintained in the recovered retroviral population. This procedure is suitable for ex pression cloning from retroviral libraries and should be applicable to the analysis of natural retrovirus populations.