Use of trypsin for rapid and efficient purification of murine sarcoma and leukemia virus.
A method is described for purification of MSV-MuLV from culture supernatant of chronically infected 78A,1 rat embryo cell line. This method involves direct polyethylene glycol-NaCl precipitation of the low speed supernatant of culture fluid followed by digestion of the pellet with trypsin. This procedure efficiently disrupts large aggregates which normally entrap most of the virus. Highly purified virus can be obtained in very good yield by a combination of sedimentation velocity and isopycnic centrifugation : yields up to 100 A280 units (17 mg of protein) of purified virus per liter of culture fluid can be observed. This procedure appears well suited for large scale isolation of virion associated enzymatic activities.
D000818:Animals; D011232:Chemical Precipitation; D017988:Gammaretrovirus / Q000302:isolation & purification*; D009052:Leukemia Virus, Murine / Q000201:enzymology / Q000302:isolation & purification*; D008722:Methods; D011092:Polyethylene Glycols / Q000032:analysis; D011494:Protein Kinases / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D051381:Rats; D009053:Sarcoma Viruses, Murine / Q000201:enzymology / Q000302:isolation & purification*; D014357:Trypsin
D011092:Polyethylene Glycols; D011494:Protein Kinases; D012194:RNA-Directed DNA Polymerase; D014357:Trypsin