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cmi12243-sup-0001-si.tif4020K

Fig. S1.R. rickettsii Adr1 mediates vitronectin acquisition and serum resistance.

A. The R. rickettsii Adr1 homologue of Adr1 is nearly identical to that of R. conorii, which had been characterized throughout this manuscript. Incubation of E. coli expressing a R. rickettsii Adr1 derivative containing an E. coli optimized signal sequence and His6-tag was incubated with NHS, followed by SDS-PAGE separation and Western blotting. IPTG-induced E. coli BL21(pRR7045) and not empty vector or uninduced controls mediated vitronectin (Vn) acquisition as demonstrated by the presence of anti-Vn bands similar to the normal human serum (NHS) control lane. R. rickettsii Adr1 expression was demonstrated through both anti- Adr1 and anti-His blots. Presence of E. coli in each lane was confirmed by anti-E. coli RNA polymerase α-subunit (RNAP).

B. Only E. coli expressing R. rickettsii Adr1 [BL21(pRR7045) + IPTG] are capable of resisting the deleterious effects of NHS. Those bacteria harbouring the empty vector (pET22b) or uninduced Adr1 plasmid [(pRR7045) − IPTG] are sensitive to NHS. Statistical comparisons were performed using the Student's t-test.

cmi12243-sup-0002-si.tif3672K

Fig. S2.R. typhi Adr1 mediates vitronectin acquisition and serum resistance.

A. R. typhi Adr1 (RT815) is 69.3% identical and 80.5% similar to R. conorii Adr1. Incubation of E. coli expressing a R. typhi Adr1 derivative containing an E. coli optimized signal sequence and His6-tag was incubated with NHS, followed by SDS-PAGE separation and Western blotting. IPTG-induced E. coli BL21(pRT815) and not empty vector or uninduced controls mediated vitronectin (Vn) acquisition as demonstrated by the presence of anti-Vn bands similar to the normal human serum (NHS) control lane. R. typhi Adr1 expression was demonstrated through anti-Adr1 blot. Presence of E. coli in each lane was confirmed by anti-E. coli RNA polymerase α-subunit (RNAP).

B. Only E. coli expressing R. typhi Adr1 [BL21(pRT815) + IPTG] are capable of resisting the deleterious effects of NHS. Those bacteria harbouring the empty vector (pET22b) or uninduced Adr1 plasmid [(pRT815) − IPTG] are sensitive to NHS. Statistical comparisons were performed using the Student's t-test.

cmi12243-sup-0003-si.tif3456K

Fig. S3.R. prowazekii Adr1 mediates vitronectin acquisition and serum resistance.

A. R. prowazekii Adr1 (RP827) is 69.5% identical and 79.9% similar to R. conorii Adr1. Incubation of E. coli expressing a R. prowazekii Adr1 derivative containing an E. coli optimized signal sequence and His6-tag was incubated with NHS, followed by SDS-PAGE separation and Western blotting. IPTG-induced E. coli BL21(pRP827) not empty vector (pET22b) or uninduced controls [(pRP827) − IPTG] mediated vitronectin (Vn) acquisition as demonstrated by the presence of anti-Vn bands similar to the normal human serum (NHS) control lane. R. prowazekii Adr1 expression was demonstrated through anti-His blot. Presence of E. coli in each lane was confirmed by anti-E. coli RNA polymerase α-subunit (RNAP).

B. Only E. coli expressing R. prowazekii Adr1 [BL21(pRP827) + IPTG] are capable of resisting the deleterious effects of NHS. Those bacteria harbouring the empty vector (pET22b) or uninduced Adr1 plasmid [(pRP827) − IPTG] are sensitive to NHS. Statistical comparisons were performed using the Student's t-test.

cmi12243-sup-0004-si.doc36KTable S1. Primers used in this study.

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