CONSTRUCTION OF YEAST EXPRESSION VECTOR CONTAINING ELT-B GENE FOR HIGH LEVEL OF HEAT LABILE ENTEROTOXIN-B SUBUNIT (LT-B)
Journal Title: International Research Journal of Pharmacy (IRJP) - Year 2011, Vol 2, Issue 7
Abstract
Enterotoxigenic Escherichia coli (ETEC) are the most common cause of diarrhea, the major disease agent of ETEC is the heat labile enterotoxin (LT-B) subunit. LT-B is a subunit vaccine candidate to be used as a strong mucosal adjuvant which enchances serum and local immune responses to co-administered antigens. In order to construct yeast expressing vector for the LT-B protein, the elt-b gene encoding LT-B was amplified from an origin E.coli DNA by PCR. The expression vector was constructed by inserting the elt-B gene into the pYES shuttle vector and recombinant vector was transformed into Saccharomyces cerevisiae and then induced its expression in the presence of galactose. The expressed protein was identified from the total soluble protein of the recombinant construct by the method of SDS – PAGE analysis. Comparative analysis were carried out to detect the quantitative of LT-B protein expressed in the construct and it was approximately 3.5% 0f the total protein. Immuno blotting showed that the maximum amount of LT-B protein expressed in yeast since the whole recombinant yeast has been considered as a new vaccine formulation the expression of LT-B in S.cerevisiae can offer an effective inexpensive strategy to protect against diarrhea at high areas.
Authors and Affiliations
Renuga G , Saravanan R , krisnan Murali , Thandapani Babu
Keywords
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