食糧 その科学と技術 No.43( )

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1 53 inclusion body: CA 2001 CA

2 54 1 Refolding on Resin Dilution additive methods

3 55 3 CA ,4- CA CA 7 CA 20 DnaJ, DnaK, GroEL/ES 1 CA 2 CA M 2

4 CA 3. CA

5 ,4- CA CA -1,4- CD CD Gellman 8 TritonX-100 -CD 7 CitSyn CD 3 -CD 17 Cyclodextrin : CD CD6 CD7 CD8 CD CD CD CD9 CD10 CD14 CD26

6 58 CA : 17 CA 9 CA CA CTAB C16 C16, C18 26 CA SDS CA CA 10 CA 3 CitSyn 49,000/ B CAB 30,000 Zn 15,000 4

7 59 S-S CitSyn 6 M 1 30 CA CD Tween40 Tween CA CA 1 5

8 60 CA CA; CA 7 CA; CA 2 B CAB 16 CitSyn CTAB SB mm DTT 6 M 16 S-S CAB CTAB SB3-14 DL- CA S-S

9 61 CA 6 1 Lowry 25 30mg/ml 6 M A S-S 40mM DTT 2 4 Tween 40, Tween 60

10 62 8 l 25 l of 8M : 6M 0.3 l of 4M DTT 40 mm (, 1 8 l 560 l of ( 0.1% 2mM DL-cystine, l of 3 % CA ( 0.6%, 1 (15,000rpm 5 min CTAB, SB CTAB SB3-14 S-S DL- S-S - :GSSG GSH 10 DL- GSH/GSSG DL- 3 3 CA CA 0.6 CA

11 63 CA CA CA CA CA CA 1 15,000rpm 5 CA 12 CA 1 SDS-PAGE

12 64 2 circular dichroism:cd BioRad BioBeads 1/ CA 5, LDL human C-type lectin like oxidized low density lipoprotein receptor hlox-1 3 C 7 hlox-1 CA 90 8 CD CA 9 hlox-1 LDL LDL LDL LDL C 12,13

13 65 C 273 C C C 183 C C 142 C N C-type lectin-like oxidized LDL receptor : hlox-1 3 S-S 3 2 : 40,000 : C-type lectin-like domain (CTLD) : 細胞外領域

14 66 1B 2B CA 1 2 CA CA 100

15 DL- S-S S-S 3 4 CA CA CA Refolding CA

16 68 Kit 1 Marston,F.A.O. 1986, Biochem.J., 240, Wetlaufer,D.B.and Xie,T 1995, Prot.Sci., 4, D.Rozema, et al., J. Am. Chem. Soc., 117, M.Yoshimoto. et al., J.Chromatogr.B, 743, Y.Akiyoshi, et al., Bioconjugate Chem., 10, R.Kuboi. et al., J.Chromatogr.B, 743, BioView, 41, D.Rozema, & S.H.Gellman., Biochemistry, 35, T.Takaha, et al., J.Biol.Chem., 271, S.Machida, et al., FEBS lett., 486, Q.Xie, et al., Protein Expression and Purification, 32, S.Machida, et al., US patent

食糧 その科学と技術 No.43( )

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