爱泼斯坦-巴尔病毒1型核抗原结合：电子显微镜观察

Epstein-Barr virus nuclear antigen type 1 binding: electron microscopy.

作者信息

Hirsch I, Reischig J, Benada O, Bartsch D, Brichácek B, Boguszaková L, Vonka V

机构信息

Department of Experimental Virology, Institute of Sera and Vaccines, Prague, Czechoslovakia.

出版信息

J Virol Methods. 1988 Dec;22(2-3):133-42. doi: 10.1016/0166-0934(88)90096-1.

DOI:10.1016/0166-0934(88)90096-1

PMID:2851598

Abstract

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.

摘要

通过DNA-琼脂糖凝胶柱层析和蓝色葡聚糖琼脂糖凝胶柱层析从Raji细胞中提取并纯化了爱泼斯坦-巴尔病毒（EBV）核抗原1型（EBNA-1）。通过电子显微镜观察其与源自EBV（M-ABA株）DNA的质粒pM765-10形成的复合物。特异性标准如下：（1）EBNA-1优先结合到pM765-10的ori-P区域；（2）用抗EBNA-1（IR-3）IgG抗体使EBV DNA/EBNA-1复合物特异性增大；（3）所得的EBV DNA/EBNA-1/抗EBNA-1抗体复合物对1.5 M NaCl处理具有抗性。形成EBV DNA/EBNA-1复合物的最佳条件是50至150 mM NaCl和pH 6.0。为了实现EBV DNA/EBNA-1复合物的高产率和特异性，EBNA-1和pM765-10需要达到平衡。