Supporting Figure 6
Fig. 6. Construction of strain BCJ118.49. Strain BCJ118.49 was constructed by two integration events into the chromosome of strain 168. In the first integration, pBCJ103.7 was integrated into the chromosome by a single crossover event through homology with the 3' end of yacF to give strain BCJ108.3. This process places lysK under the endogenous lysS expression signals while removing the lysS structural gene from the transcriptional control of its own promoter and transcriptionally isolating it between terminators (lollipop structures). The lysS gene is partially removed in the second integration event, a double crossover event between the vector sequences and the 3' end of the lysS gene of linearized pBCJ113.15, giving strain BCJ118.49.
