69
Views
0
CrossRef citations to date
0
Altmetric
Original Articles

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

, , , , , , & show all
Pages 46-56 | Published online: 17 Dec 2009
 

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.

ACKNOWLEDGMENTS

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

Reprints and Corporate Permissions

Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?

To request a reprint or corporate permissions for this article, please click on the relevant link below:

Academic Permissions

Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?

Obtain permissions instantly via Rightslink by clicking on the button below:

If you are unable to obtain permissions via Rightslink, please complete and submit this Permissions form. For more information, please visit our Permissions help page.