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. 2003 Dec 1;31(23):6996–7002. doi: 10.1093/nar/gkg883

Figure 1.

Figure 1

Construction of E.coli strains in which the rpsA TIRs from various γ-proteobacteria govern translation of the chromosomal lacZ gene. The PCR fragments carrying the rpsA regulatory regions have been first cloned in-phase with the α-peptide gene (‘lacZ’) of pEMBLΔ46, generating pES1γ (pES1Sty, pES1Yp, pES1Baph, etc.), and then transferred onto the chromosome of the Lac– E.coli strain (ENSO) by homologous recombination between lac sequences present on both the plasmid and chromosome. ENSO carries a deletion encompassing the lac promoter/operator region and ribosome-binding site of the lacZ gene. Plac and op are the promoter and operator of the lac operon.