A plasmid which overproduces the Tet(M) resistance protein (pSH52)

Value Proposition

Bacterial resistance to current therapies remains a clinical challenge. For example, numerous enterococcal, staphylococcal, and streptococcal bacterial species are known to exhibit resistance to potent antibacterial tetracycline through upregulation of a ribosomal protection protein called Tet(M). Model systems which allow for the in vitro investigation of this protein, including mechanisms of gene expression, are needed to develop new therapies which can combat antibacterial resistance.

Technology

This invention is a plasmid which contains the tetracycline resistance gene Tet(M) under direct control of a potent promoter. Specifically, it includes the streptococcal transposon Tn916, which contains the Tet(M) gene, under control of the T7 phage 10 promoter. Thus, expression of the gene in vitro requires the addition of the T7 RNA polymerase. In one version of the plasmid, an inducible T7 RNA polymerase is also included under control of a lac promoter, which can be induced in the in vitro system using IPTG.

Advantages

• This plasmid contains the Tet(M) gene under direct control of a potent promoter
• This plasmid can also contain inducible RNA polymerase
• This plasmid allows for the direct introduction and inducible expression of Tet(M) in vitro