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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제19권 제5호
- 발행연도
- 2009.1
- 수록면
- 502 - 510 (9page)
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초록· 키워드
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Although the Escherichia coli heat-labile enterotoxin B
subunit (LTB) has already been expressed in several
different systems, including prokaryotic and eukaryotic
organisms, studies regarding the synthesis of LTB into
oligomeric structures of pentameric size in the budding
yeast Saccharomyces cerevisiae have been limited. Therefore,
this study used a functional signal peptide of the amylase
1A protein from rice to direct the yeast-expressed LTB
towards the endoplasmic reticulum to oligomerize with
the expected pentameric size. The expression and assembly
of the recombinant LTB were confirmed in both the cellfree
extract and culture media of the recombinant strain
using a Western blot analysis. The binding of the LTB
pentamers to intestinal epithelial cell membrane glycolipid
receptors was further verified using a GM1-ganglioside
enzyme-linked immunosorbent assay (GM1-ELISA). On
the basis of the GM1-ELISA results, pentameric LTB
proteins comprised approximately 0.5-2.0% of the total
soluble proteins, and the maximum quantity of secreted
LTB was estimated to be 3 mg/l after a 3-day cultivation
period. Consequently, the synthesis of LTB monomers
and their assembly into biologically active oligomers in a
recombinant S. cerevisiae strain demonstrated the feasibility
of using a GRAS microorganism-based adjuvant, as well
as the development of carriers against mucosal disease.
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참고문헌 (22)
참고문헌 신청
[
Arakawa, T. / 1997 / Expression of cholera toxin B subunit oligomers in transgenic potato plants / Transgenic Res 6 : 403 ~ 413
Bradford, M. M / 1976 / A rapid and sensitive method for the quantification of microgram quantities of protein using the principle of protein-dye binding / Anal. Biochem 72 :
Brake, A. J. / 1984 / Alpha-factor-directed synthesis and secretion of mature foreign proteins in Saccharomyces cerevisiae / Proc. Natl. Acad. Sci. U.S.A. 81 : 4642 ~ 4646
De Geus, B. / 1997 / A comparison of natural and recombinant cholera toxin B subunit as stimulatory factors in intranasal immunization / Vaccine 15 : 1110 ~ 1113
Arakawa, T. / 1997 / Expression of cholera toxin B subunit oligomers in transgenic potato plants / Transgenic Res 6 : 403 ~ 413
Bradford, M. M / 1976 / A rapid and sensitive method for the quantification of microgram quantities of protein using the principle of protein-dye binding / Anal. Biochem 72 :
Brake, A. J. / 1984 / Alpha-factor-directed synthesis and secretion of mature foreign proteins in Saccharomyces cerevisiae / Proc. Natl. Acad. Sci. U.S.A. 81 : 4642 ~ 4646
De Geus, B. / 1997 / A comparison of natural and recombinant cholera toxin B subunit as stimulatory factors in intranasal immunization / Vaccine 15 : 1110 ~ 1113
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