The pTriEx<TMSYMBOL></TMSYMBOL>-4 vector is uniquely designed to allow rapid characterization of target genes in multiple expression systems. With this vector a single recombinant plasmid can be used to test expression in E. coli, insect and vertebrate cells. Transient vertebrate expression is mediated by the CMV immediate early enhancer and promoter2. For expression in insect cells, pTriEx-4 contains flanking baculovirus sequences to permit the generation of recombinant baculoviruses using the BacVector<TMSYMBOL></TMSYMBOL> System. In baculovirus-infected insect cells, expression is driven by the very late p10 promoter. Expression in E. coli is regulated by the tightly controlled T7lac promoter. Expression can be induced in hosts such as NovaBlue by infecting with λCE6, a phage that constitutively expresses T7 RNA polymerase from the λpL and λlpI promoters. Alternatively, pTriEx recombinant plasmids can be transferred into a (DE3)pLacI host that allows IPTG based induction. Native protein can be expressed by cloning into the NcoI site, or native protein can be generated by cloning into the PshAI or SmaI sites and cleavage of the fusion protein with Enterokinase or Thrombin, respectively. The pTriEx-4 vector is also available as an Ek/LIC Cloning Kit.
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