An improved vector for the expression of proteins in all three translational reading frames

Abstract

We have constructed a novel vector (pN-7) that is capable of producing large amounts of recombinant proteins in E. coli and requires minimal manipulation for the construction of recombinant expression vectors. This expression vector (pN-7) contains the tightly regulated λ pL promoter, cII ribosome binding site, and initiator condon ATG. The pN-7 vector also contains cleavage sites for the restriction enzymes SmaI, EcoRV, and HpaI that provide blunt ends in all three reading frames. Thus after cleavage with the appropriate restriction enzyme, this novel vector can be directly ligated to the DNA fragment that contains the open reading frame without further manipulation.