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. 2009 Mar 12;2009:520283. doi: 10.1155/2009/520283

Figure 1.

Figure 1

Generation and characterization of ibeR deletion mutant from the meningitic strain E44. (a) Generation of the ibeR deletion mutant (BR2). Two DNA fragments flanking ibeR were amplified and ligated into the suicide vector pCVD442 to construct pCBR2. The ibeR deletion mutants were generated through gene allele exchanges and suicide vector loss. The arrowheads indicate primer locations. (b) Verification of the ibeR deletion by PCR. The parent strain E44 and the ibeR deletion mutant BR2 were verified though colony PCR with primers listed in Table 2. (c) Invasion of HBMEC with the E. coli parent E44 carrying pWKS30, the ibeR mutant BR2 with pWKS30, and the complemented BR2 carrying the ibeR locus in pWKS30. E44/pWKS, BR2/Pwks, and BR2/pWKS1030 were incubated with HBMEC monolayers and the standard invasion assay was carried out as described in Section 2. The results are expressed as relative invasion%. Columns marked with ∗∗ are significantly different (P < .01).