Journal: BMC Microbiology

Article Title: PgFur participates differentially in expression of virulence factors in more virulent A7436 and less virulent ATCC 33277 Porphyromonas gingivalis strains

doi: 10.1186/s12866-019-1511-x

Figure Lengend Snippet: Analysis of expression of selected glycosyltransferase gene and glycosylation pattern of glycoconjugates produced by P. gingivalis wild-type (A7436 and ATCC 33277) and pgfur mutant (TO6 and TO16) strains. Bacteria were grown under high- (Hm) or low-iron/heme (DIP) conditions in liquid culture medium. a Relative gene expression was determined at the indicated time points in pgfur mutant (TO6 and TO16) strains versus wild-type (A7436 and ATCC 33277) strains by RT-qPCR. b Whole bacterial lysates were separated by SDS-PAGE, transferred onto nitrocellulose membranes and probed with selected lectins. MAL II, sialic acid; AAL, fructose linked to N -acetylglucosamine with α-1,6 linkage or to N -acetyllactosamine with α-1,3 linkage, arabinose; SNA, sialic acid linked to the terminal galactose with α-2,6 or α-2,3 linkage; RCA 120, galactose and N -acetylgalactosamine; JACALIN, galactose linked to N -acetylgalactosamine with β-1,3 linkage. Equal loading of proteins was visualized in SDS-PAGE gels by staining with Coomassie Brilliant Blue G-250 (loading control). * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: After blocking with Carbo-Free Blocking Solution (Vector Laboratories), membranes were incubated with biotinylated lectins (1:3300; Vector Laboratories) in 50 mM Tris–HCl, pH 7.5, containing 150 mM NaCl (TBS), 0.2% Tween-20, 1 mM CaCl 2 and 1 mM MnCl 2 .

Techniques: Expressing, Produced, Mutagenesis, Quantitative RT-PCR, SDS Page, Staining