Cloning and analysis of the glucosyl transferase gene encoding type I antigen in Shigella flexneri.
The O-antigen of most Shigella flexneri serotypes contains an identical tetrasaccharide repeating unit. Apart from serotype Y, the O-antigen is modified by addition of a glucosyl and/or O-acetyl residue to a specific position in the O-unit. In this study the glucosyl transferase gene from a serotype 1 a has been cloned and identified. The bacteriophage SfV integrase (int) gene was used to probe a S. flexneri Y53 (serotype 1 a) cosmid library and 18 unique clones were identified. Southern hybridisation of these clones indicated two unlinked regions of the chromosome contained the int homologue. When expressed in a live candidate vaccine strain of S. flexneri serotype Y (SFL124), clones with one region produced type I antigen, whereas clones containing the other region produced mainly type Y antigen. One of the cosmid clones positive for type I antigen by agglutination and Western blotting was selected for further study. Genes involved in O-antigen glucosyl modification were mapped on a 5.8 kb fragment and subclones were produced which fully or partially expressed the type I antigen, depending on the extent of the clone. Fully and partially expressing clones may be useful vaccine candidate strains for protection against disease caused by two serotypes of S. flexneri.
Bastin
D A
DA
Division of Biochemistry and Molecular Biology, School of Life Sciences, Faculty of Science, Australian National University, Canberra, Australia.
Lord
A
A
Verma
N K
NK
D000942:Antigens, Bacterial / Q000737:chemistry / Q000235:genetics*; D002240:Carbohydrate Sequence; D003001:Cloning, Molecular; D003360:Cosmids; D005798:Genes, Bacterial; D005964:Glucosyltransferases / Q000235:genetics*; D006031:Glycosylation; D019426:Integrases / Q000235:genetics; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D012763:Shigella flexneri / Q000201:enzymology / Q000235:genetics* / Q000276:immunology*
D000942:Antigens, Bacterial; D005964:Glucosyltransferases; D019426:Integrases