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. 2010 Sep 1;41(3):729–740. doi: 10.1590/S1517-83822010000300025

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Figure 1. — Primer design for construction of genedeleted mutants and the recombinant plasmid pGMB151(fljB::lacZ).

Specific primer pairs (P1A/B, P2A/B) located up- and down-stream of flhDC and fliA were designed to amplify homologic fragments that were then linked as the gene defective recombinant fragments. Primer pair P1A/B specific to the sequence up- and downstream of ompR were designed to amplify the homologic fragment, which was digested by NspV and BssHII and linked as the ompR defective recombinant fragment.

Primer pairs (P1A/B, P2A/B) specific to the sequence up- and downstream of fljB were designed to amplify two homologic fragments that were directionally linked with the promoterless lacZ cassette as the recombinant fragment. Recombinant fragments were individually cloned into the suicide plasmids for construction of gene-deleted mutants and the recombinant plasmid pGMB151(fljB::lacZ).