Murine Type-C Virus Group-Specific Antigens: Interstrain Immunochemical, Biophysical, and Amino Acid Sequence Differences

Abstract

MATERIALS AND METHODS Murine leukemia virus (MuLV). Several strains of mouse leukemia virus were used in these studies. Rauscher virus (R-MuLV) was grown in monolayer cultures of chronically infected mouse (BALB/c) bone marrow, JLS-V9 cells (50). The virus-shedding cell line was obtained from Electro-Nucleonics, Bethesda, Md. AKR virus was obtained from NIH Swiss embryo cells infected with an isolate from the spleen of an AKR mouse. These cells were kindly provided by Janet Hartley (National Cancer Institute). The La Puente isolate (1504E) of the wild mouse leukemia virus (WMLV) was obtained from Earle Officer (University of Southern California, Los Angeles, Calif.) (31; M.

Downstream processing of cell culture-derived virus particles

“…The latter is based on the buoyant density of the particle and the rate zonal separation on the particle size and mass. Examples of virus particles that were purified by gradient centrifugation techniques are feline leukemia virus [48], Gibbon ape lymphoma virus [49], hepatitis B virus [50], influenza virus [51], Japanese encephalitis virus [52], mammalian type C virus [53], mouse mammary tumor virus [54], mouse oncorna virus [55], mumps virus [56], murine leukemia virus [57], tick-borne encephalitis virus [58], turkey coronavirus [25], rabies virus [59] and Vaccinia virus [60]. An alternative centrifugation method is the differential centrifugation where pelleting (Epstein-Barr virus [61] and hepatitis A virus [62]) or simple sedimentation onto a sucrose cushion (Moloney murine sarcoma virus [63]) is done once or repeatedly at different centrifugation speeds.…”

Immunologic studies of the low molecular weight DNA binding protein of murine oncornaviruses

“…Serial bleedings were also obtained once every two weeks after the last injection. Double immunodiffusion was done in plates that contained 0.8 % agarose, PH 7.4, ionic strength 0.15 according to Oroszlan et al (1974). Plates were observed over '7 A POLYMERASE BINDING -m* 50 1w I" a 72-h period at room temperature and photographed when precipitation lines were maximally developed.…”

Solubilized tsta and the major viral structural proteins, gp70 and p30, in the immune response to murine leukemias induced by friend and rauscher virus

“…gp70 and p30 were purified from Rauscher virus as described previously (August and Strand, 1973 ;Oroszlan et al, 1974) and stored at -20". Batches were tested for antigenicity by binding to goat antisera before use.…”

“…The structural proteins p30 and gp70 from Rauscher virus used in these studies were highly purified as judged by a variety of criteria (August et al, 1973;Oroszlan et al, 1974). First, they were homogeneous by several biochemical criteria including SDS electrophoresis and, in the case of p30, sequencing of the amino terminal amino acids.…”