Molecular analysis of endogenous virus ev21-slow feathering complex of chickens. 1. Cloning of proviral-cell junction fragment and unoccupied integration site.

Sex-linked slow-feathering gene, K, is genetically associated with the presence of an avian endogenous retrovirus ev21 in White Leghorns (WL). An EcoRI fragment corresponding to the endogenous virus ev21-cell junction fragment and a fragment homologous to the proviral unoccupied site (US) were cloned, respectively, from genomic DNA libraries of two WL chickens: an ev21-only female and an ev-negative male. A 1.7-kilobase pairs (kbp) fragment cleaved from the cloned proviral US by the HaeIII restriction endonuclease was the most informative probe to molecularly characterize the occupied and unoccupied integration sites of ev21 locus. Restriction fragment length polymorphism analysis of slow-feathering (SF) and rapid-feathering (RF) chickens from various commercial breeds, using the US HaeIII 1.7-kbp probe, indicated that the complete genetic association between ev21 and SF phenotype is common among other lines of SF chickens and was not restricted to WL. It was also shown that there was at least one additional DNA region highly homologous to DNA sequences flanking the EV21 integration site in the chicken genome. In SF birds of either sex this additional repeat was distinguishable from the site occupied by ev21 (OR) and represents an unoccupied repeat (UR). Analysis of DNA from RF revertant females showed novel patterns of reversion. In Type I RF revertants, RF is associated with the complete excision of proviral ev21 DNA sequences. In Type II revertants, the UR homologous to the cell sequences flanking ev21 integration site is excised, but proviral ev21 sequences remain intact. A hypothesis to explain these types of reversion is suggested. It postulates a close association between OR and UR on the Z chromosome.