Method for reproducible large-volume production and purification of Rauscher murine leukemia virus.
Rauscher murine leukemia virus was produced in roller-bottle cultures of chronically infected JLS-V9 cells. Virus from this culture fluid was concentrated and purified by two semi-isopycnic bandings in sucrose gradients. Virus material obtained from young, nonconfluent cultures (early-harvest virus) yielded products characteristically containing endogenous ribonucleic acid-dependent deoxyribonucleic acid polymerase with high specific activity (400 to 1,000 pmol of [3H]thymidine 5'-triphosphate incorporated per milligram of protein per hour). Fluids obtained from older confluent cultures (late-harvest virus) yielded products with endogenous ribonucleic acid-dependent deoxyribonucleic acid polymerase with little or no specific activity (200 pmol or less of [3H]thymidine 5'-triphosphate incorporated per milligram of protein per hour), but with higher virus particle counts and greater amounts of protein and gs antigen than the early-harvest products.
D002460:Cell Line; D002474:Cell-Free System; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D011925:Rauscher Virus / Q000254:growth & development* / Q000276:immunology / Q000302:isolation & purification; D013942:Thymine Nucleotides / Q000378:metabolism; D014764:Viral Proteins / Q000096:biosynthesis; D014776:Virus Cultivation / Q000379:methods*; D014779:Virus Replication
D013942:Thymine Nucleotides; D014764:Viral Proteins; D012194:RNA-Directed DNA Polymerase