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dc.creatorVrat Kamboj,Dev
dc.creatorNema,Vijay
dc.creatorKumar Pandey,Arun
dc.creatorKumar Goel,Ajay
dc.creatorSingh,Lokendra
dc.date2006-10-01
dc.date.accessioned2019-05-03T12:44:22Z
dc.date.available2019-05-03T12:44:22Z
dc.identifierhttps://scielo.conicyt.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500010
dc.identifier.urihttp://revistaschilenas.uchile.cl/handle/2250/84915
dc.descriptionStaphylococcal food poisoning (SFP) is caused by the members of superantigen family called staphylococcal enterotoxins (SEs). About 20 different types of SEs are produced by Staphylococcus aureus out of which type A (SEA), B (SEB), C (SEC) and D (SED) are commonly implicated in SFP. Among these, SEB is the most potent toxin and has also gained the status of biological warfare (BW) agent. Therefore, detection of SEB is of utmost importance. Any immunological detection system for SEB requires specific and sensitive antibodies which inturn depends on the purity of the SEB. In the present investigation, seb gene of S. aureus was cloned and expressed in E. coli along with biotin as fusion partner to facilitate the purification process. The yield of purified recombinant SEB was 13.1 mg/L of culture broth. Biotin tag from the biotinylated toxin was removed by protease cleavage, and both biotinylated and non-biotinylated toxin types were used for raising hyperimmune antiserum. Antisera were also specific for SEB amongst different kinds of food poisoning agents tested by indirect plate ELISA and western blot analysis. The quality of the antisera raised in this study was found superior to the commercially available antiserum. The investigation suggests that construction of recombinant staphylococcal enterotoxin B is a good alternative for production of pure enterotoxin to be used in antibody generation.
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dc.languageen
dc.publisherPontificia Universidad Católica de Valparaíso
dc.relation10.4067/S0717-34582006000500010
dc.rightsinfo:eu-repo/semantics/openAccess
dc.sourceElectronic Journal of Biotechnology v.9 n.5 2006
dc.subjectaffinity purification
dc.subjectbiotin fusion
dc.subjectELISA
dc.subjectgene cloning
dc.subjectSEB
dc.subjectwestern blot
dc.titleHeterologous expression of staphylococcal enterotoxin B (seb) gene for antibody production


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