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Recombinant production of Epstein-Barr virus BZLF1 trans-activator and characterization of its DNA-binding specificity

Protein Expr Purif. 2015; 
Lim CS, Goh SL, Krishnan G, Ng CC.
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Codon Optimization … Positive clones were identified by colony PCR, using BZLF1 forward primer and T7 reverse primer (a pET102 vector primer … because (i) codon adaptation index (CAI) for native BZLF1 is 0.61, whereas 1.0 is the ideal CAI (Rare Codon Analysis Tools: http://www.genscript.com/cgi … Get A Quote

摘要

This paper describes the recombinant production of a biologically active Epstein-Barr virus BZLF1 trans-activator, i.e., Z-encoded broadly reactive activator (ZEBRA), that recognized specific DNA motifs. We used auto-induction for histidine-tagged BZLF1 expression in Escherichia coli and immobilized cobalt affinity membrane chromatography for protein purification under native conditions. We obtained the purified BZLF1 at a yield of 5.4mg per gram of wet weight cells at 75% purity, in which 27% of the recombinant BZLF1 remained biologically active. The recombinant BZLF1 bound to oligonucleotides containing ZEBRA response elements, either AP-1 or ZIIIB, but not a ZIIIB mutant. The recombinant BZLF1 showed a speci... More

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