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. 2011 Jul 7;7(7):e1002117. doi: 10.1371/journal.ppat.1002117

Figure 8. Analysis of invA expression in Y. enterocolitica O:3.

Figure 8

(A) An overview of the invA promoter region including the IS1667 insertion of Y. enterocolitica O:3 strains is shown. The transcriptional start sites of the invA gene and from the predicted IS1667-encoded promoter are indicated by broken arrows, the dark boxes indicate the RovA binding sites identified in the homologous invA promoter of Y. pseudotuberculosis. The thick line represents the invA promoter sequence and the thin line illustrates the IS1667 sequence. The arrow indicates the gene encoding the putative transposase of the IS1667 element. Sites used for the upstream deletion constructs are indicated by arrows. The numbers indicate the position of the deletion relative to the transcriptional start site of the invA gene. (B) Overnight cultures of Y. enterocolitica O:3 strain Y1 harbouring the PinvA O:8::luxCDABE (pFU170), PinvA O:3::luxCDABE (pFU171), PinvA O:3ΔIS::luxCDABE (pFU172) and PIS1667::luxCDABE (pFU202) fusion constructs were diluted (1∶100) and grown in LB at 37°C for four hours and luciferase activity was determined. (C) Expression by progressive deletion of the invA 5′-regulatory region was analyzed in Y. enterocolitica O:3 Y1 and the isogenic rovA mutant derivative Y12 harbouring the PinvAO:3::luxCDABE fusion. The numbers indicate the 5′ end points of the regulatory region of invA from Y. enterocolitica O:3 in the fusion constructs relative to the transcriptional start site (+1). The luciferase activity determined from the cultures is given in relative light units (RLU) and represents the mean ± standard deviation of at least three independent experiments. (D) Sequence of the 3′-end of the IS1667 inserted into invA of Y. enterocolitica O:3 at position −143 is shown. The −10 and −35 region of the predicted IS1667-encoded promoter are indicated. Sites used for the upstream deletion constructs are indicated by arrows. The numbers indicate the position of the deletion relative to the transcriptional start site of the invA gene.