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Figure 3 | BMC Biochemistry

Figure 3

From: Topological characterisation and identification of critical domains within glucosyltransferase IV (GtrIV) of Shigella flexneri

Figure 3

Functional analysis of deletion constructs. a) LPS Western blots and slide agglutination results. Deletion constructs were transformed into a serotype Y strain (SFL1616) to test for modification of O-antigen to serotype 4a. SFL 2324 is GtrIV fused to PhoA/LacZ and then transformed into SFL1616. The bright band is indicative of the serotype conversion from serotype Y to serotype 4a. The rest of the deletions show absence of modification to serotype 4a when probed with Type IV antisera. Results of the slide agglutination assays are indicated below the Western blots. A score of +ve indicates a positive result while a score of -ve indicates a negative result. b) Western blots (using anti-alkaline phosphatase primary antibody) performed on the membrane protein extractions of the various GtrIV loop deletion constructs that were fused to alkaline phosphatase. B2347 and B2348 were not detected, indicating GtrIV Loop No. 2 and Loop No. 3 deletions have a direct effect on the assembly of the protein in the cellular membrane. B2349, B2350 and B2352 were detected in lower amounts, the amount of protein that is assembled in the cellular membrane in these three deletion mutants is much lower as compared to the fully intact protein (B2339). The intense band seen for B2351 is almost the same as the fully intact protein.

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