Replication and protein expression in a range of bacterial hosts.
- Expression vector is engineered for high-copy replication and transcription in a variety of bacteria hosts.
- Vector backbone offers a multicloning site and the versatile Streptococcus feacalis kanamycin resistance gene for selection in gram-negative and grampositive bacteria.
Many contemporary molecular biological approaches are predicated on the ability to express a foreign protein of interest in a bacterial host. Protein expression, however, must often be screened through several different bacterial hosts to identify one that will fold the protein properly. Unique origins of replication and cis-acting promoter elements in each host complicate matters, requiring the investigator to invest significant time to transfer the gene-of-interest into in new expression vector for each new host. To address this inadequacy, Emory inventors have developed a single universal vector that can express a protein of interest in a variety of different bacterial hosts. With a modified origin of replication, a broad host promoter, and the kanamycin resistance gene from Streptococcus feacalis the vector can be replicated, transcribed, and selected in range of different gram-negative and gram-positive bacteria. An inexpensive, quick-to-market strategy that includes packaging the universal vector with a series of testable bacterial strains would fill an important need in the basic research community.
Proof-of-principle experiments indicate empty vector replicates in Esherichia coli, Borrelia burgdorferi, Acinetobacter baylyi, and Streptococcus pyogenes; and the lacZ reporter gene is expressed in E. coli and A. baylyi, creating blue colonies on media containing X-gal. Other industrial stains are being tested.