Supplementary data
Global transcriptional analysis of acid-inducible genes in Streptococcus mutans: multiple two-component systems involved in acid adaptation, by Y. Gong, X.-L. Tian, T. Sutherland, G. Sisson, J. Mai, J. Ling and Y.-H. Li
Microbiology vol. 155, part 10, pp. 3322-3332
Fig. S1. Strategy for construction and confirmation of gene deletion mutants by PCR ligation mutagenesis. The example shown here is the SmciaH mutant (see text for the more detailed description of the method). The ligation product was transformed into S. mutans UA159 to generate a ciaH deletion mutant SmciaH (ΔciaH::Em, Ermr) by allelic replacement through double crossover recombination. The mutant was genetically confirmed by PCR using a combination of the primers shown. The PCR profiles of the mutant and the parent were compared to determine successful construction. The anticipated fragments and sizes of PCR products are presented in the gel image. Note: the PCR products of ciaH-P1-Erm-P2 (1681 bp) and Erm-P1-ciaH-P4 (1511 bp) were obtained only if the ligation product was inserted into the correct location and in the correct orientation. Lane 1, 1 kb ladder. Lanes 2&endash;5 (mutants): ciaH-P1-Erm-P2 fragment, 1681 bp; Erm-P1-ciaH-P4 fragment, 1511 bp; ciaH-P1-P4 fragment, 2332 bp; Erm-P1-P2 fragment, 860 bp. Lanes 6&endash;9 (parent): PCR-generated negative results for the parent strain, except for primer pair ciaH-P1-P4 that generated an anticipated 2614 bp fragment. The fragments of ciaH-P1-Erm-P2 and Erm-P1-ciaH-P4 from the mutant were sequenced to confirm the location and orientation of the insertion.
Table S1. Primers used for qRT-PCR and construction of the mutants in this study.
Table S2. Genes differentially expressed in S. mutans during acid adaptation.
Copyright © 2009 Society for General Microbiology.
