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. 2010 Jan 29;192(7):1975–1987. doi: 10.1128/JB.01474-09

FIG. 4.

FIG. 4.

Phenotype of fimS-exchanged strains. (A) CBB-stained SDS-12.5% PAGE gel (upper) and Western blot (lower) with anti-type I FimA antibody. Total protein samples were denatured at 80°C for 20 min before SDS-PAGE. Lane 1, 33277; 2, AGFS1; 3, AGFS1(pTCBex); 4, AGFS1(pTCBex33277fimS); 5, AGFS1(pTCBexW83fimS); 6, W83; 7, WFS1; 8, WFS1(pTCBex); 9, WFS1(pTCBex33277fimS); 10, WFS1(pTCBexW83fimS). The CBB-stained single band of W83 FimA in lane 9 is indicated by a thin arrow. A thick arrow indicates the signals from 41-kDa monomers. (B) CBB-stained SDS-12.5% PAGE gel (upper) and Western blot with anti-W83 FimA antibody (lower). Total protein samples were denatured at 80°C for 20 min before being subjected to SDS-PAGE. Lane 1, W83; 2, W83(pTCBex); 3, W83(pTCBex33277fimS); 4, W83(pTCBexW83fimS); 5, WFS1; 6, WFS1(pTCBex); 7, WFS1(pTCBex33277fimS); 8, WFS1(pTCBexW83fimS); 9, 33277. Stained bands corresponding to W83 FimA in lanes 3 and 7 are indicated by an arrow. (C) RT-PCR confirming the expression of fimS in the complemented fimS knockout mutants. The 500-bp fimS cDNAs were amplified with the primer set shown in Fig. 1D and Table 1. After 25 cycles of PCR, an equal volume of each reaction mixture was fractionated by 3% AGE. The glucose kinase gene (glk; 415 bp) was coamplified in each reaction as an internal control. Lane 1, 33277; 2, AGFS1(pTCBex); 3, AGFS1(pTCBex33277fimS); 4, AGFS1(pTCBexW83fimS); 5, W83; 6, W83(pTCBex); 7, W83(pTCBex33277fimS); 8, W83(pTCBexW83fimS); 9, WFS1(pTCBex); 10, WFS1(pTCBex33277fimS); 11, WFS1(pTCBexW83fimS).