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Figure 3 | BMC Biotechnology

Figure 3

From: Differences in virulence of pneumolysin and autolysin mutants constructed by insertion duplication mutagenesis and in-frame deletion in Streptococcus pneumoniae

Figure 3

Construction of the ply and lytA knockout mutants by IFD. In step 1, linear DNA fragments containing flanking regions of the ply (A) and lytA (B) genes, generated by PCR, were transformed into IDM-ply (A) and -lytA (B), respectively, for a second round of homologous recombination. The second recombination event could result in either restoration of the original gene, or mutants of IFD-ply (A) and -lytA (B) with a double-crossover. IFD-ply (A) and -lytA (B) mutants were selected by decreasing concentration of erythromycin (erm) for ply or chloramphenicol (cat) for lytA, and increasing concentrations of ampicillin. Primer sets ply-p1/ply-P2 or lytA-P1/lytA-P2 were used to verify the IFD mutants.

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BMC Biotechnology

ISSN: 1472-6750

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