ThepMyvector series: A versatile cloning platform for the recombinant production of mycobacterial proteins inMycobacterium smegmatis

Structural and biophysical characterization of molecular mechanisms of disease-causing pathogens, such asMycobacterium tuberculosis, often requires recombinant expression of large amounts highly pure protein. For the production of mycobacterial proteins, overexpression in the fast-growing and non-pathogenic speciesMycobacterium smegmatishas several benefits over the standardEscherichia coliexpression strains. However, unlike forE. coli, the range of expression vectors currently available is limited. Here we describe the development of the pMy vector series, a set of expression plasmids for recombinant production of single proteins and protein complexes inM. smegmatis. By incorporating an alternative selection marker, we show that these plasmids can also be used for co-expression studies. All vectors in the pMy vector series are available in the Addgene repository ().