Comparison of JC and BK human papovaviruses with simian virus 40: restriction endonuclease digestion and gel electrophoresis of resultant fragments.
JC virus was found to have a buoyant density of 1.20 g/cm(3) in linear sucrose-D(2)O and 1.35 g/cm(3) in cesium chloride isopycnic gradients. DNA extracted either from JC-infected cultures or from gradient-purified virions occupied a dense position relative to linear DNA in cesium chloride/ethidium bromide gradients, and the circular configuration of the extracted DNA was confirmed by electron microscopy, with a measured molecular weight of 2.93 x 10(6). DNA from BK virus was similarly prepared and compared to JC and to an SV40 DNA standard by digestion with restriction endonuclease preparations from Haemophilus influenzae, Haemophilus parainfluenzae, and Escherichia coli. Digests were electrophoretically analyzed on gradient polyacrylamide slab gels or agarose gels, and the three viruses were found to have distinctly different cleavage patterns by this form of analysis: JC and BK viruses were almost entirely different from SV40 and significantly different from each other. Thus, JC and BK human papovaviruses appear to be discrete new members of the papovavirus group, rather than SV40 variants.
D002499:Centrifugation, Density Gradient; D004270:DNA, Circular / Q000032:analysis* / Q000302:isolation & purification; D004279:DNA, Viral / Q000032:analysis* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004720:Endonucleases / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D006190:Haemophilus / Q000201:enzymology; D006193:Haemophilus influenzae / Q000201:enzymology; D008854:Microscopy, Electron; D008970:Molecular Weight; D027383:Papillomaviridae / Q000032:analysis* / Q000145:classification; D027382:Polyomaviridae; D011120:Polyomavirus / Q000032:analysis*; D013539:Simian virus 40 / Q000032:analysis*; D013936:Thymidine; D014316:Tritium
D004270:DNA, Circular; D004279:DNA, Viral; D014316:Tritium; D004720:Endonucleases; D013936:Thymidine