Expression of Shiga toxin B subunit at cell surface in E. coli K-12.
The three parts (Stx17B, Stx27B and StxB) of Shiga toxin B subunit have been fused into a cell surface exposed loop of the LamB protein at a BamH I site between residues 153 and 154. Western blotting revealed that the three parts of Shiga toxin B subunit could be expressed as the LamB fusion proteins in E. coli. Indirect immunofluorescence and immunoelectron microscopy analyses showed fusion proteins LamB/Stx17B and LamB/Stx27B could be expressed at cell surface in E. coli, but fusion protein LamB/StxB could not be expressed at cell surface; it was aggregated in cytoplasm and was toxic to host. This expression system provided a new way to construct an oral live vaccine against Shigella dysenteriae 1.
Su
G F
GF
Institute of Biotechnology, Academy of Military Medical Sciences, CPLA, Beijing, PRC.
Brahmbhatt
H N
HN
Timmis
K N
KN
D001425:Bacterial Outer Membrane Proteins; D001427:Bacterial Toxins / Q000096:biosynthesis / Q000235:genetics*; D010582:Bacteriophage lambda / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D018272:Porins; D011991:Receptors, Virus / Q000096:biosynthesis / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D022603:Shiga Toxins; D012762:Shigella dysenteriae
D001425:Bacterial Outer Membrane Proteins; D001427:Bacterial Toxins; D018272:Porins; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins; D022603:Shiga Toxins; C016532:maltoporins