Segregation, viral phenotype, and proviral structure of 23 avian leukosis virus inserts in the germ line of chickens

We have artificially introduced 23 avian leukosis virus (ALV) proviral inserts into the chicken germ line by injection of wild-type and recombinant subgroup A ALV near the blastoderm of fertile eggs just before incubation. Eight viremic males were identified as germline mosaics because they transmitted proviral DNA to their generation 1 (G-1) progeny at a low frequency. Eleven female and 9 male G-1 progeny carried 23 distinct proviruses that had typical major clonal proviral-host DNA junction fragments detectable after digestion of their DNA with SacI, Southern blotting and hybridization with a probe representing the complete ALV genome. These proviruses, identified by their typical proviral-host DNA junction fragments, were transmitted to approximately 50% of their G-2 progeny after mating the G-1 parents to a line of chickens lacking endogenous ALV proviral inserts. One G-1 female carried 2 proviruses and another 3. The proviruses appeared to be scattered throughout the genome. One of the 14 proviruses carried by females was on the sex (Z) chromosome. Two of the 3 proviruses carried by a single G-1 female were linked with a recombination frequency of about 0.20. Twenty-one of the proviruses coded for infectious ALV. Two proviruses coded for envelope glycoprotein, and cell cultures carrying them were relatively resistant to subgroup A sarcoma virus, but failed to produce infectious ALV. One of these proviruses coded for internal gag proteins, had a deletion in pol, but produced non-infectious virus particles. The other failed to code for gag proteins and had no detectable internal deletions nor did it produce virus particles. Thus, we have shown that replication-competent ALV can artificially infect germ-line cells and that spontaneous defects in the inherited proviruses occur at a rather low rate.