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. 2021 Apr 13;6:82. [Version 1] doi: 10.12688/wellcomeopenres.16665.1

Figure 4. Cloning strategy for the construction of the C. trachomatis/E. coli individual transposon and transposase shuttle vectors pSW2-mCh-C9-ΔTpase and pSW2-mCh-C9-ΔTpon.

Figure 4.

A: Deleting the active site of the C9 transposase from pSW2-mCh-C9. The active site of the C9 transposase in pSW2-mCh-C9 was deleted by PCR using primers 13 and 14 ( Table 1), adding MreI sites to both ends. The resulting PCR product was re-ligated to form pSW2-mCh-C9-ΔTpase. This deletion (426 bp) is designed to inactivate the transposase generating a truncated protein of predicted molecular weight 17.4kDa ( Figure 5C). The scissor symbol indicates the location of the deletion. B: Deletion of the transposon unit from pSW2-mCh-C9. The transposon unit was deleted from pSW2-mCh-C9 by PCR using primers 11 and 12 ( Table 1), adding MreI sites to both ends. The resulting PCR product was re-ligated to form pSW2-mCh-C9-ΔTpon. The scissor symbol indicates the location of the deletion.