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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2010.1
- 수록면
- 319 - 324 (6page)
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초록· 키워드
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Experimental bioinformatics data obtained from an E. coli cell-based eukaryotic protein purification experiment were analyzed in order to identify any bottleneck as well as the factors affecting the target purification. All targets were expressed as His-tagged maltose-binding protein (MBP) fusion constructs and were initially purified by immobilized metal affinity chromatography (IMAC). The targets were subsequently separated from the His-tagged MBP through TEV protease cleavage followed by a second IMAC isolation. Of the 743 total purification trials, 342 yielded more than 3 mg of target proteins for structural studies. The major reason for failure of target purification was poor TEV proteolysis. The overall success rate for target purification decreased linearly as cysteine content or isoelectric point (pI) of the target increased. This pattern of pI versus overall success rate strongly suggests that pI should be incorporated into target scoring criteria with a threshold value.
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참고문헌 (24)
참고문헌 신청
Altschul, S. F. / 1990 / Lipman DJ: Basic local alignment search tool / J. Mol. Biol. 215 : 403 ~ 410
Bessette, P. H. / 1999 / Efficient folding of proteins with multiple disulfide bonds in the Escherichia coli cytoplasm / Proc. Natl. Acad. Sci. U.S.A. : 13703 ~ 13708
Chrunyk, B. A. / 1993 / Inclusion body formation and protein stability in sequence variants of interleukin-1 beta / J. Biol. Chem. 268 : 18053 ~ 18061
Dirk Schneider / 2006 / High Level Expression of a Protein Precursor for Functional Studies / BMB reports 39 (6) : 717 ~ 721
Fahnert, B. / 2004 / Murine Wnt-1 with an internal c-myc tag recombinantly produced in Escherichia coli can induce intracellular signaling of the canonical Wnt pathway in eu
Bessette, P. H. / 1999 / Efficient folding of proteins with multiple disulfide bonds in the Escherichia coli cytoplasm / Proc. Natl. Acad. Sci. U.S.A. : 13703 ~ 13708
Chrunyk, B. A. / 1993 / Inclusion body formation and protein stability in sequence variants of interleukin-1 beta / J. Biol. Chem. 268 : 18053 ~ 18061
Dirk Schneider / 2006 / High Level Expression of a Protein Precursor for Functional Studies / BMB reports 39 (6) : 717 ~ 721
Fahnert, B. / 2004 / Murine Wnt-1 with an internal c-myc tag recombinantly produced in Escherichia coli can induce intracellular signaling of the canonical Wnt pathway in eu
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