Fig. 3

Immunoblot of recombinantly expressed S. pneumoniae capsule in E. coli. Lysed, whole cell samples were separated by SDS–PAGE on a Bolt 4–12% bis–tris gel run with MOPS buffer. Glycan was detected using anti-serotype 4 primary antibody and anti-rabbit fluorescent secondary antibody. M molecular weight marker PageRuler Plus. Presence or absence of pB4 recombinant S. pneumoniae serotype 4 capsule is denoted below the lanes as ± respectively, as are the strain names. a lpxM deletion. b wecA deletion and insertion of gne. Presence or absence of gne is denoted below the lanes as ± respectively;  + ^P denotes gne is provided on pMAF12 plasmid,  + ^C denotes chromosomal integration of gne in place of wzzE-wecA. c waaL deletion. The presence or absence of waaL and wecA are denoted below the lanes as ± respectively. In the case of Hobby strain gne is also integrated on the chromosome. d Chain length modification. The presence or absence of wzzB and wzDE are denoted below the lanes as ± respectively. In the case of WzDE + ^P denotes the presence of pEXT21:wzDE and + ^C indicated that wzDE are integrated in place of wzzB on the chromosome