Molecular cloning of the avian myelocytomatosis virus genome and recovery of infectious virus by transfection of chicken cells

Abstract

The avian retrovirus myelocytomatosis virus 19 (MCV) possesses an interesting diversity of oncogenic potentials but the virus has proven difficult to study because of its inability to replicate without the assistance of a helper [myelocytomatosis-associated] virus. The genome of MCV was isolated and amplified by molecular cloning in a prokaryotic [phage .lambda.gtWES.cntdot..lambda.B] vector. The topography of the cloned DNA was explored by the use of restriction endonucleases and radioactive complementary DNA representing specific domains in avian retrovirus genomes. The cloned DNA appeared to be an authentic representation of the MCV genome: the size and genetic topography of the DNA were comparable to those of MCV and transfection of the cloned DNA into chicken embryo fibroblast cells (in company with the DNA of a suitable helper virus) gave rise to virus with the genome and transforming potentials of MCV. The avilability of cloned MCV DNA should facilitate a variety of genetic and biochemical manipulations directed at elucidating the mechanism of oncogenesis by MCV.