Molecular cloning of the avian erythroblastosis virus genome and recovery of oncogenic virus by transfection of chicken cells.

摘要: Avian erythroblastosis virus (AEV) causes and sarcomas in birds transforms both erythroblasts fibroblasts to neoplastic phenotypes culture. The viral genetic locus required for oncogenesis by AEV is at present poorly defined; moreover, we know very little of the mechanism tumorigenesis virus. To facilitate further analysis these problems, used molecular cloning isolate genome as recombinant DNA a procaryotic vector. identity isolated was verified mapping with restriction endonucleases tests biological activity. circular form unintegrated purified from synchronously infected quail cells cloned into EcoRI site lambda gtWES x B. A endonuclease cleavage map established. By hybridization complementary probes representing specific parts avian retrovirus genomes, DNAs correlated map. These data show that nucleotide sequences unique comprise least 50% are located approximately middle genome. Our confirm extend previous descriptions obtained other procedures. We studied detail two clones containing DNA: topography virtually identical, except one clone apparently contained copies terminal redundancy occurs linear cells; probably only copy redundant sequence. recover infectious DNA, developed procedure transfection compensated defectiveness replication. accomplished this ligating (Rous-associated type 1) whose could complement deficiencies AEV. Ligation facilitated using neutral fragment linker between otherwise noncompatible termini. Cloned gave rise capable transforming bone marrow culture inducing erythroleukemia chickens. conclude represent authentic undisturbed procedure. Molecular offers powerful approach identification characterization genomes.