High-level expression of the Epstein-Barr virus alkaline deoxyribonuclease using a recombinant baculovirus: application to the diagnosis of nasopharyngeal carcinoma.
The Epstein-Barr virus (EBV) alkaline deoxyribonuclease (DNase) was inserted into the baculovirus Autographa californica nuclear polyhedrosis virus (AcMNPV). Infection of the insect cell line Spodoptera frugiperda (SF9) with the recombinant virus led to the expression of an enzymatically active alkaline DNase. The recombinant EBV alkaline DNase was highly soluble, and the recombinant baculovirus produced approximately 10-20 mg of EBV DNase per 1 X 10(9) cells. The recombinant enzyme activity was neutralized by specific antisera to the EBV DNase and was recognized by these sera in Western blot analysis and immunofluorescence tests. The recombinant EBV DNase was neutralized by these sera from patients with nasopharyngeal carcinoma and chronic infectious mononucleosis. Western blot analysis using these patients' sera showed that IgG and IgA antibodies to the EBV DNase could be readily detected.
Baylis
S A
SA
Cancer Research Campaign Laboratories, Paterson Institute for Cancer Research, Christie Hospital and Holt Radium Institute, Withington, Manchester, United Kingdom.
Purifoy
D J
DJ
Littler
E
E
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D015153:Blotting, Western; D002460:Cell Line; D003851:Deoxyribonucleases / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D004854:Herpesvirus 4, Human / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight; D009303:Nasopharyngeal Neoplasms / Q000175:diagnosis* / Q000382:microbiology; D010957:Plasmids; D014162:Transfection
D003851:Deoxyribonucleases