FIGURE 1.

Properties of the pETAB cassette for regulated expression of MccE492. Genes mceAB were cloned under a T7 promoter with further regulation by lac operator (A). A consensus ribosome binding site (RBS) was included upstream of each coding region. This cassette was introduced in the BamHI site of p33AM, generating p33pETAB (B) that expresses constitutively lacI^q, to repress expression from the T7 promoter in the absence of IPTG. Different versions of the mceA genes can be replaced in this construction using the NdeI/HindIII restriction fragment.