034 12.0   463 6.5 25.58 0.107, 0.007 15.4   354 5.9 34.62 0.004, 0.207 47.9   465 6.3 25.49 0.077, 0.006 13.2   381 5.6 29.61 0.003, 0.032 10.4   491 7.0 22.69 0.356, 0.012 29.8   406 6.2 28.54 0.006, 0.081 14.0   494 6.5 23.16 1.400, 0.062 22.8   418 6.3 27.97 0.089, 1.927 21.6   495 6.7 23.13 6.875, 0.025 278.1 Outer surface protein C (GI:3914248) 452 6.6 26.33 0.006, 0.246 42.5 30S ribosomal R428 datasheet protein S4 (B7J2H5) Phosphoglycolate phosphatase

(GI:226320487), and hypothetical (GI:226315606) 497 6.3 22.87 0.262, 0.022 12.1   479 6.4 24.51 0.060, 0.648 10.7   519 7.1 20.08 0.734, 0.027 26.8   501 6.5 22.47 0.030, 1.956 64.6 Same as 505 525 6.4 21.03 0.234, 0.008 30.9 Neutrophil activating protein (GI:15595035) 505 6.3 22.33 0.017, 0.570 34.0 OspC (GI: 226246807) Neutrophil activating protein (GI:15595035) 528 6.2 20.95 0.068, 0.004 15.9   517 6.2 21.41 0.002, 0.095 54.4   541 6.4 20.31 0.097, 0.005 20.8   543 5.6 19.67 0.006, 0.072 11.9 Rapamycin solubility dmso   559 5.6 17.70 0.137, 0.008 17.2   551 6.2 19.51 0.075, 0.762 10.2   581 4.9 11.91 2.069, 0.048 42.7 6.6 kDa lipoprotein (GI:1477781) 573 5.3 14.07 0.005, 0.255 55.0   585 6.3 28.02 0.125, 0.010 12.2               586 6.1 44.19 0.357, 0.001 674.8 *Flagellin (GI:120230)             587 6.1 44.41 0.209, 0.000 765.7               588 6.1

41.54 0.276, 0.001 527.4               * Flagellin appears to be produced at equivalent levels in both strains but fold change depicted is higher for respective spots due to slight mobility differences of this protein in B31 and N40D10/E9 gels. To further evaluate the differences in the proteins that are differentially expressed in the two strains, a limited MALDI mass spectrometric (MALDI-MS) analysis of selected protein spots was conducted. The proteins identified by MALDI-MS are listed in Table 1. Interestingly,

three protein spots of slightly different mobility, number 586 in B31 and numbers 272 and 293 in N40D10/E9, were found to be more abundant (>650 times) than Myosin that of the equivalent spots in the compared strain. Our MALDI-MS analysis (Table 1) identified them as flagellin proteins. We amplified the flagellin gene (bb0147) from B31 and N40D10/E9 strains and sequenced the PCR product from the N40 strain. Sequence analysis showed a single amino acid change resulting in slight difference in the pI of the two proteins. This could affect mobility of the flagellin of each strain slightly on a 2D gel with each appearing as more abundant protein relative to the other B.