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| Studying on Bio-characteristics of Expression Product of Shiga-like Toxin IIe A Subunit gene and ELISA Test |
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Abstract In order to get the soluble expression products, Shiga-like toxin IIe A subunit was analyzed by TMpred software. Then a pair of primers were designed and the expression plasmid pGEX-A was constructed before being transformed into BL21 (DE3). SDS-PAGE and Western blotting results showed the gene fragment was expressed abundant soluble protein and the expression products had immunogenicity. The assay of it’s bio-characteristic toxicity showed that the soluble fusion protein lost the toxicity as SLT-IIe. Polyclonal antiserum produced by immunizing the New Zealand rabbits with the purified expression product restrained the toxicity of SLT-IIe to Vero E6 cells because it had significantly high neutralizing activity (90.4℅) against SLT-IIe. But the protective efficacy test showed that the immunization with the expression products could not reduce mortality of the mice. The ELISA method was established using the expressed protein to detect anti-SLT-IIe A IgG. The assay had excellent specificity, sensitivity and reduplication and could be applied to detect the edema disease in piglets.
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Received: 09 October 2006
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