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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제28권 제10호
- 발행연도
- 2018.1
- 수록면
- 1,749 - 1,759 (11page)
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초록· 키워드
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Recombinant (rec) prion protein (PrP) is an extremely useful resource for studying protein misfolding and subsequent protein aggregation events. Here, we report mass production of high-purity rec-polypeptide encoding the C-terminal globular domain of PrP; (90-230) for human and (89-231) for murine PrP. These proteins were expressed as His-tagged fusion proteins in E. coli cultured by a high cell-density aerobic fermentation method. RecPrPs recovered from inclusion bodies were slowly refolded under reducing conditions. Purification was performed by a sequence of metal-affinity, cation-exchange, and reverse-phase chromatography. The current procedure yielded several dozens of milligrams of recPrP per liter with >95% purity. The purified recPrPs predominantly adopted an α-helix-rich conformation and were functionally sufficient as substrates to measure the seeding activity of human and animal prions. Establishment of a procedure for high-level production of highpurity recPrP supports the advancement of in vitro investigations of PrP including diagnosis for prion diseases.
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참고문헌 (56)
참고문헌 신청
Prusiner SB / 1998 / Prions / Proc. Natl. Acad. Sci. USA 95:13363~13383
Pan K-M / 1993 / Conversion of a-helices into b-sheets features in the formation of the scrapie prion proteins / Proc. Natl. Acad. Sci. USA 90:10962~10966
Cohen FE / 1998 / Pathologic conformations of prion proteins / Annu. Rev. Biochem 67:793~819
Caughey B / 2009 / Getting a grip on prions: oligomers, amyloids, and pathological membrane interactions / Annu. Rev. Biochem 78:177~204
Prusiner SB / 1982 / Novel proteinaceous infectious particles cause scrapie / Science 216:136~144
Pan K-M / 1993 / Conversion of a-helices into b-sheets features in the formation of the scrapie prion proteins / Proc. Natl. Acad. Sci. USA 90:10962~10966
Cohen FE / 1998 / Pathologic conformations of prion proteins / Annu. Rev. Biochem 67:793~819
Caughey B / 2009 / Getting a grip on prions: oligomers, amyloids, and pathological membrane interactions / Annu. Rev. Biochem 78:177~204
Prusiner SB / 1982 / Novel proteinaceous infectious particles cause scrapie / Science 216:136~144
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