Purification of simian immunodeficiency virus, SIVMAC251, and of its external envelope glycoprotein, gp148.
Two-phase extraction in a system composed of dextran and polyethylene glycol was used to purify simian immunodeficiency virus, SIVMAC251 (32H isolate) from 25 l of culture supernatant. The virus partitioned to the interphase with 80% recovery of gag peptide p27 and reverse transcriptase and an about 25% recovery of the external env glycoprotein, gp148. The virus was treated with octylglycoside and its subcomponents separated. Two gag-p27 containing fractions were obtained; gag-1, which also contained reverse transcriptase and nucleopeptides, and gag-2, which contained the major portion of the p27. The env gp148 was purified by chromatography through a series of lectin columns. The prepared materials are characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and immuno- and lectin blotting.
Gilljam
G
G
Department of Clinical Virology, Swedish Institute for Infectious Disease Control, Solna.
Siridewa
K
K
Hammar
L
L
D015153:Blotting, Western; D002240:Carbohydrate Sequence; D002460:Cell Line; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D015686:Gene Products, env / Q000302:isolation & purification*; D015683:Gene Products, gag / Q000302:isolation & purification; D006023:Glycoproteins / Q000302:isolation & purification*; D006801:Humans; D037102:Lectins; D008969:Molecular Sequence Data; D015302:Simian Immunodeficiency Virus / Q000302:isolation & purification* / Q000378:metabolism
D015686:Gene Products, env; D015683:Gene Products, gag; D006023:Glycoproteins; D037102:Lectins