CONSTRUCTION AND EXPRESSION OF A SPLICED VARIANT OF EPSTEIN-BARR VIRUS BZLF1 AND PREPARATION OF ITS POLYCLONAL ANTIBODY

Abstract

The BZLF1 gene-encoded protein, Zta (EB1, ZEBRA), is a key transcriptional activator of induction of the lytic cycle of EBV. Zta; it contains a basic region with homology to the DNA binding domains of the AP-1 family. In this study, an alternatively spliced BZLF1 (ΔBZLF1) cDNA lacking exon 2, which encodes the DNA-binding domain of Zta, was isolated from B95-8 marmoset cell line releasing EBV. The cDNA was inserted into a prokaryotic expression vector pET-28a+. The His-tagged recombinant protein was overproduced in E. coli BL21(DE3) and purified by nickel affinity chromatography. The purified fraction was characterized by Western blot and MALDI-TOF-MS analysis and used as an antigen to immunize mice. The antibody against ΔZta can recognize both denatured and natural Zta protein. The ΔZta protein and its antibody can be used to further investigate its unknown functions.

Keywords:

• antibody
• BZLF1
• purification
• spliced variant
• Zta

ACKNOWLEDGMENTS

This work was supported by National Basic Research Program of China (973 Program) (2006CB504305) and Beijing Natural Science Foundation (No. 7051006).