Jellyfish venom C-CFTX1-STxB chimeric antigen subcloning, expression and its antigenicity assay in laboratory mouse

Box jellyfish inflicts painful stings and may be lifethreatening. The venom of C. fleckeri contains a variety of bioactive proteins that are cytolytic, cytotoxic, inflammatory or lethal. Two of the most abundant proteins contained in the nematocysts of C. fleckeri; CfTX-1 and CfTX-2 were difficult to separate using electrophoretic or chromatographic methodsRecombinant expression technology may offer an alternative to the isolation of native C. fleckeri venom protein. The aim of this study is to undertake novel expression studies of the cytolytic C.fleckeri toxin C-CfTX1-STxB in E.coli and study its antigenicity in Syrian mice.cftx1 construct based on bioinformatics designed and synthetic gene prepared in plasmid pUC57. 576 bp C-cftx1 cloned with PCR and was subcloned in a pET28a-stxB expression vector with BamHI and SalI restriction enzyme sites and transformed into E.coli BL21 (DE3) competent cells. C-cftx1-stxB gene expression was artificially induced by IPTG, and protein purified from the gel. Also, Mice were challenged by the Rhopilema nomadica s venom. In this experimental study, C-CfTX1-STxB 822 bp gene was confirmed by PCR, sequencing and enzymatic analysis. Besides Recombinant protein was confirmed by SDS-PAGE and Western blotting. Serum was prepared from immunized mice after blood sampling, and the produced antibody was quantitated by ELISA. The results showed that immunized mice in a challenge after 60 days tolerated 50x LD50 of jellyfish venom. Considering that recombinant C-CfTX1-STxB protein doesn t have cardiotoxicity and neurotoxicity effects on mice, this produced protein can be suggested as a jellyfish venom vaccine candidate for mice or at a later stage of a clinical trial for humans.

کلیدواژه ها:

Chironex fleckeri ، Jellyfish venom ، C-CFTX1-STxB chimeric antigen ، Antigenicity assay