Epstein-Barr virus nuclear antigen type 1 binding: electron microscopy.
Epstein-Barr virus (EBV) nuclear antigen type-1 (EBNA-1) was extracted and purified from Raji cells by chromatography on DNA-Sepharose and Blue-dextran Sepharose. Its complexes with plasmid pM765-10 derived from EBV (strain M-ABA) DNA were visualized by electron microscopy. The criteria of specificity were as follows: (1) preferential binding of EBNA-1 to the ori-P region of pM765-10; (2) specific enlargement of EBV DNA/EBNA-1 complexes with anti-EBNA-1 (IR-3) IgG antibody; and (3) resistance of the resulting EBV DNA/EBNA-1/anti-EBNA-1 antibody complexes to treatment with 1.5 M NaCl. The optimal conditions for the formation of EBV DNA/EBNA-1 complexes were 50 to 150 mM NaCl and pH 6.0. A balanced equilibrium of EBNA-1 and pM765-10 was necessary to achieve both a high yield and specificity of EBV DNA/EBNA-1 complexes.
Hirsch
I
I
Department of Experimental Virology, Institute of Sera and Vaccines, Prague, Czechoslovakia.
Reischig
J
J
Benada
O
O
Bartsch
D
D
Brichácek
B
B
Boguszaková
L
L
Vonka
V
V
D000956:Antigens, Viral / Q000302:isolation & purification*; D001665:Binding Sites; D002460:Cell Line; D002467:Cell Nucleus / Q000276:immunology; D004279:DNA, Viral / Q000302:isolation & purification / Q000648:ultrastructure; D019309:Epstein-Barr Virus Nuclear Antigens; D004854:Herpesvirus 4, Human / Q000276:immunology* / Q000648:ultrastructure; D006801:Humans; D008854:Microscopy, Electron; D010957:Plasmids
D000956:Antigens, Viral; D004279:DNA, Viral; D019309:Epstein-Barr Virus Nuclear Antigens