Figure 6. Cloning strategy for the construction of the C. trachomatis/E. coli transposon shuttle vector pSW2-RSGFP-Tpon.

Primers 15 and 16 ( Table 1) were used to amplify the transposon unit carrying the β-lactamase gene (yellow) from plasmid pRPF215-Bla. These primers introduce a PfoI restriction site at both ends of the PCR product. This was then cloned into pRSGFP-5KO-ΔBla via its unique PfoI site to generate plasmid pSW2-RSGFP-Tpon.