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Mutagenesis Services> | . The wild-type construct of Zta (bZIP DNA binding domain (DBD) plus 50 flanking amino acids) was obtained as a N-terminal GST construct cloned into a modified pDEST15 MAGIC vector [19]. Mutant constructs of Zta (C189S, C189T, C189V, and C189A) were generated via site-directed mutagenesis of the wild-type construct (GenScript). | Get A Quote |
Zta is a bZIP transcription factor (TF) in the Epstein-Barr virus that binds unmethylated and methylated DNA sequences. Substitution of cysteine 189 of Zta to serine (Zta(C189S)) results in a virus that is unable to execute the lytic cycle, which was attributed to a change in binding to methylated DNA sequences. To learn more about the role of this position in defining sequence-specific DNA binding, we mutated cysteine 189 to four other amino acids, producing Zta(C189S), Zta(C189T), Zta(C189A), and Zta(C189V) mutants. Zta and mutants were used in protein binding microarray (PBM) experiments to evaluate sequence-specific DNA binding to four types of double-stranded DNA (dsDNA): 1) with cytosine in both strands (... More