원문정보
초록
영어
Escherichia coli heat-labile enterotoxin B subunit (LTB) is known not only to induce strong immune responses but also to be an efficient mucosal adjuvant towards either co-administrated unrelated antigens or tolerogenic T-cell epitopes. Although it has already been expressed in several different systems including prokaryotic as well as eukaryotic organisms, studies on the synthesis of the LTB into oligomeric structure of pentameric size in budding yeast Saccharomyces cerevisiae has been limited. In these studies, with a functional signal peptide of
amylase 1A protein from rice, the yeast-expressed LTB was targeted to endoplasmic reticulum to oligomerize with the expected size of pentamer. The xpression and assembly of recombinant TB was observed in both cell-free extract as well as culture filtrate of recombinant strain by Western blot analysis. The binding of LTB pentamers to intestinal epithelial cell membrane glycolipid receptors was further confirmed by GM1-ganglioside enzyme-linked immunosorbent
assay (GM1-ELISA). Based on the results of GM1-ELISA, pentameric LTB proteins comprise approximately 0.1 - 0.2% of total soluble protein and the maximum amount of secreted LTB was estimated to be 100mg/L after the 3-day cultivation.