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

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Abstract
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.