Open AccessAlternative System for Detection and Mapping of Activation Domains
Author(s) -
Srdjan Ašković,
R Baumann
Publication year - 1997
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/97225rr02
Subject(s) - lytic cycle , biology , acetyltransferase , chloramphenicol acetyltransferase , gene , reporter gene , open reading frame , heterologous , activator (genetics) , gene expression , fusion protein , herpes simplex virus , transcription (linguistics) , regulation of gene expression , heterologous expression , genetics , microbiology and biotechnology , computational biology , virus , recombinant dna , acetylation , peptide sequence , linguistics , philosophy
The ZEBRA protein is a transcriptional activator that induces expression of viral lytic genes in cells harboring latent Epstein-Barr virus (EBV). In this report it is shown that a derivative of ZEBRA that cannot activate transcription (Zd) can be used to detect and characterize activation domains. Three expression vectors that allow the fusion of putative activation regions in any reading frame were constructed using Zd. These vectors were used to demonstrate the activity of different classes of activation domains using a chloramphenicol acetyltransferase (cat) reporter gene construct containing seven ZEBRA response elements (Z7). The Zd/Z7 system effectively detected proline-rich, glutamine-rich and acidic activation domains in a variety of cell lines and cell types. Using a bioassay unique to the EBV Zd/Z7 system, fusion constructs can also be tested for the ability to activate gene expression directly from a chromatin structure, the EBV genome. These studies indicate that the Zd/Z7 system is an alternative to GAL4 and can be a useful tool for identifying heterologous activation domains.