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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제20권 제11호
- 발행연도
- 2010.1
- 수록면
- 1,563 - 1,570 (8page)
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초록· 키워드
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To construct a highly sensitive detection system for endocrine disruptors (EDs), we have compared the activity of promoters with the n-alkane-inducible cytochrome P450gene (ALK1), isocitrate lyase gene (ICL1), ribosomal protein S7 gene (RPS7), and the translation elongation factor-1α gene (TEF1) for the heterologous gene in Yarrowia lipolytica. The promoters were introduced into the upstream of the lacZ or hERa reporter genes,respectively, and the activity was evaluated by β-galactosidase assay for lacZ and Western blot analysis for hERα. The expression analysis revealed that the ALK1and ICL1 promoters were induced by n-decane and by EtOH, respectively. The constitutive promoter of RPS7and TEF1 showed mostly a high level of expression in the presence of glucose and glycerol, respectively. In particular, the TEF1 promoter showed the highest β-galactosidase activity and a significant signal by Western blotting with the anti-estrogen receptor, compared with the other promoters. Moreover, the detection system was constructed with promoters linked to the upstream of the expression vector for the hERα gene transformed into the Y. lipolytica with a chromosome-integrated lacZ reporter gene under the control of estrogen response elements (EREs). It was indicated that a combination of pTEF1phERαand CXAU1-2XERE was the most effective system for the E2-dependent induction of the β-galactosidase activity. This system showed the highest β-galactosidase activity at 10-6 M E2, and the activity could be detected at even the concentration of 10-10 M E2. As a result, we have constructed a strongly sensitive detection system with Y.
lipolitica to evaluate recognized/suspected ED chemicals,such as natural/synthetic hormones, pesticides, and commercial chemicals. The results demonstrate the utility,sensitivity, and reproducibility of the system for identifying and characterizing environmental estrogens.
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참고문헌 (23)
참고문헌 신청
[학술지(정
Arnold, S. F / 1996 / A yeast estrogen screen for examining the relative exposure of cells to natural and xenoestrogens / Environ. Health Perspect 104 : 544 ~ 548
Barth, G / 1993 / Cloning of the isocitrate lyase gene (ICL1) from Yarrowia lipolytica and characterization of the deduced protein / Mol. Gen. Genet 241 : 422 ~ 430
Barth, G / 1997 / Physiology and genetics of the dimorphic fungus Yarrowia lipolytica / FEMS Microbiol. Rev 19 : 219 ~ 237
Bradley, M / 2001 / Polarizability fields for use in three-dimensional quantitative structure-activity relationship (3D-QSAR) / J. Chem. Inf. Comput. Sci 41 : 1301 ~ 1307
Arnold, S. F / 1996 / A yeast estrogen screen for examining the relative exposure of cells to natural and xenoestrogens / Environ. Health Perspect 104 : 544 ~ 548
Barth, G / 1993 / Cloning of the isocitrate lyase gene (ICL1) from Yarrowia lipolytica and characterization of the deduced protein / Mol. Gen. Genet 241 : 422 ~ 430
Barth, G / 1997 / Physiology and genetics of the dimorphic fungus Yarrowia lipolytica / FEMS Microbiol. Rev 19 : 219 ~ 237
Bradley, M / 2001 / Polarizability fields for use in three-dimensional quantitative structure-activity relationship (3D-QSAR) / J. Chem. Inf. Comput. Sci 41 : 1301 ~ 1307
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