[33] High-expression vectors with multiple cloning sites for construction of trpE fusion genes: pATH vectors

This chapter describes the structure and applications of a series of Escheriehia coli plasmids, the pATH plasmids, designed for the production of proteins from any cloned DNA sequence that contains an open reading frame (ORF). The cloned DNA sequences are fused in-frame to the trpE gene of E. coli, which codes for anthranilate synthase. Thus, the hybrid protein produced contains the amino-terminal 323 residues ofanthranilate synthase followed by the translation product specified by the cloned DNA. The advantages of pATH plasmids for production of hybrid proteins are as follows: (1) pATH plasmids are maintained in E. coli at high copy number. (2) The plasmids are relatively small, and their complete sequence is known. (3) Multiple cloning sites (MCS) are present following codon 323 of the trpE gene, for easy construction of in-frame gcne fusions. Different pATH plasmids carry MCS in each of the three registers of the translational reading frame.