1 5 3 30 70 2/3 3 1
2 1 10 V 1 1 4 E 1 G gustducin G PCR RT-PCR G 1992 G gustducin 2 gustducin gustducin 1996 3 RT-PCR 1992 gustducin 1993 G 4 1999 2 T1r1, T1r2 TR1, TR2 5 T1r1 T1r2 T1r
3
4 T1rT1r1, T1r2 2 T1r1 T1r2 4 Sac 6 Sac T1rSac Sac T1r T1r PCR RT-PCR 7 T1r1, T1r2 T1r T1r3 T1r3 2 T1r T1r3 T1r1, T1r2 T1r3 1 4 Sac 3 2 Sac Thr55Ala, Ile60Thr, Pro61Leu, Arg371Glu, Ile706Thr 4 T1r3 Sac 2001 T1r3 5T1r3 8,9,10,11,12 Sac G T1r3 129/SvC57BL/6 T1r3 C57BL/6T1r3 11 T1r3 T1r2 T1r3
5 Tlr1 Tlr2 Tlr3 T1r2 11 T1r3 T1r1 13 T1r3 T1r1 L- 14 1 T1r 2001 T1r3 2000 15 PROP 5 T1r1 T1r2 T2r TR1 TR2 T1r1 T1r2 T1r T2r 40 80 T2r 5 7 12 T2r Cyx Qui Soa Rua Glb 6 Prp Prp T2r
6 12 9 T2r Prp 6 T2r T2r 6 mt2r5 Cyx 16 T2r Cyx PROP Qui Soa T2r T2r T2r T2r T2r 17 1 2 G gustducin T1r 1 T1r3 T1r2 2T1r3 T1r2T2r 3 T1r1 T1r3 T1r1 4 G gustducin T2r gustducin 3 4 gustducin
7
8 18 T1r1 T1r3 T1r3 T1r1 5 gustducin T2rT1r3 T1r2 6 gustducin T1r gustducin 7 T1r3 T1r2 T1r1 T1r1 T1r2 T1rT2r T1r3 2 19, 20 T1r3
9 T1r3 2 db/db db/db 8 9 21 2000
10 10 22, 23 24
11 10 2 BDNF Brain-Derived Neutrophic Factor; BDNF trkb Neurorophin Receptor Tyrosine Kinases B; B 25 BDNF BDNF 1 2 3 Sonic hedgehog Shh Shh Patched1 Ptc 26 10 Ptc Shh Ptc Ptc Shh Shh Ptc Shh Ptc
12
13 Mash1 27 11 Mash1 gustducin Mash1NeuroD 28 Mash1 Shh Nkx2.212 Shh 29 1 Hiji, Y.: Selective elimination of taste responses to sugars by proteolytic enxymes. Nature, 256, 427-429 1975 2 McLaughlin, S.K., McKinnon, P.J. and Margolskee, R.F.: Gustducin is a tastecell-specific G protein closely related to the transducins. Nature, 357, 563-569 1992
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15 15 Adler, E., Hoon, M.A., Mueller, K.L., Chandrashekar, J., Ryba, N.J.P. and Zuker, C.S.: Novel Family of Mammalian Taste Receptors. Cell, 100, 693-702 2000 16 Chandrashekar,,J., Mueller, K.L., Hoon, M.A., Adler, E., Feng L., Guo, W., Zuker, C.S. and Ryba, N.J.P: T2rs Function as Bitter Taste Receptors.: Cell, 100, 703-711, 2000 17 Caicedo, A. and Roper, S.D.: Taste Receptor Cells That Discriminate Between Bitter Stimuli. Scinece, 291, 1557-1760 2001 18 Kim, M.-R., Kusakabe, Y., Miura, H., Shindo, Y., Ninomiya, Y. and Hino, A.: Regional expression pattern of taste receptors and gustducin in the mouse tonge. Biochem. Biophys. Res. Commun., 312, 500-506 2003 19 Damak, S., Rong, M., Yasumatsu, K., Kokrashvili, Z., Varadarajan, V., Zou, S., Jiang, P., Ninomiya, Y. and Margolskee, R.F.: Detection of sweet and umami taste in the absence of taste receptor T1r3. Science, 301, 850-853 2003 20 Zhao, G.Q., Zhang, Y., Hoon, M.A., Chandrashekar,.J, Erlenbach, I., Ryba, N.J. and Zuker, C.S.: The receptors for mammalian sweet and umami taste. Cell, 115, 255-266 2003 21 Kawai, K., Sugimoto, K., Nakashima, K., Miura, H. and Ninomiya, Y.: Leptin as a modulator of sweet taste sensitivities in mice. Proc. Natl. Acad. Sci. USA., 97, 11044-11049 2000 22 Beidler, L.M. and Smallman, R.L.: Renewal of cells within taste buds. J. Cell. Biol., 27, 263-272 1965 23 Farbman, A.I.: Renewal of taste bud cells in rat circumvallate papillae. Cell Tissue. Kinet., 13, 349-357 1980 24 Stone, L.M., Finger, T.E., Tam, P.P. and Tan, S.S.: Taste receptor cells arise from local epithelium, not neurogenic ectoderm. Proc. Natl. Acad. Sci. USA, 92, 1916-1920 1995 25 Nosrat, C.A., Blomlof, J., ElShamy, W.M., Ernfors, P. and Olson, L.: Lingual deficits in BDNF and NT3 mutant mice leading to gustatory and somatosensory disturbances, respectively. Development 124, 1333-1342 1997 26 Miura, H., Kusakabe, Y., Sugiyama, C., Kawamatsu, M., Ninomiya, Y., Motoyama, J. and Hino, A.: Shh and Ptc are associated with taste bud maintenance in the adult mouse. Mech. Dev., 106, 143-145 2001 27 Kusakabe, Y., Miura, H., Hashimoto, R., Sugiyama, C., Ninomiya, Y. and Hino, A.: The neural differentiation gene Mash-1 has a distinct pattern of expression from the taste reception-related genes gustducin and T1R2 in the taste buds. Chem. Senses, 27, 445-451 2002
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17 DNA SEM, Scanning electron microscope TEM, transmission electron microscope X NMR AFM, atomic force microscopyspm, scanning probe microscopy 1 SPM SPM AFMSNOMNSOM, Scanning near-field optical microscope SPM 1980 SPM
18 SPM SPM 1981 SPM STM, Scanning tunneling microscope 1986AFM FFM, Friction force microscope MFM, Magnetic force microscopekelvin KPM, Kelvin probe microscopespm SNOM 1984 SPM SPM AFM SNOM SPM SPM STM Scanning tunneling microscope AFM Atomic force microscope FFM Friction force microscope VEM Visco-elasticity microscope MFM Magnetic force microscope Kelvin KFM Kelvin force microscope SThM (Scanning thermal microscope SNOM Scanning near-field optical microscope
19 SPM AFM SPM 10 m 20nm 0.1nm 10.5nm AFM SNOM SPM SNOM SNOM/ShFM, Scanning near-field optical / shear force microscope SNOM/AFM, Scanning near-field optical / atomic force microscope SNOM/AFM AFM 1 SNOM SNOM/AFM SNOM/ShFM 1 SNOM/AFM AFM 1 a
20 50nm 1 b nm SNOM/ AFM50nm SNOM/AFM SPM AFM 2
21 SPMm m m 1 m SNOM/AFM SPM SNOM/AFM
22 AFM AFM DNA 2 3 4 AFM AFM 3 a 40-50 nm 3 b DNA 30nm NaCl 2.0 M NaCl 5 AFM AFM AFM SEM 6 AFM
23 AFM 4 AFM 7, 8 1 m 4 nm 4 AFM AFM 9 5 a 5 b 0.38 MPa 5 c 0.48 MPa 5 d
24
25 AFM 1 m 50nm AFM AFM 10 AFMSEM DNA FISH Fluorescence in situ hybridization FISH DNA SNOM/AFM DNA DNA 6
26 ea47 15Alexsa532 532 nm 570nmPNA peptide nucleic acid DNA 2 DNA YOYO-1 488 nm 510 nm DNA Methyltrimethoxysilane 11 12 488532 nm APD avalanche photodiode6a DNA Alexa532 6b 300nm SNOM/AFM 15 13, 14 DNA 15 DNA 16 DNA SNOM/AFM nm SNOM/AFM 17
27 SNOM/AFM 7 7 a SNOM/AFM 7 b c DNA FISH 8 a FISH 1 1 AFM SNOM/AFM AFM 8 b, c 8 b AFM nm SNOM/AFM 10AFM 8 c 8 b AFM SNOM/AFM 1 2
28 18, 19 SNOM/AFM FISH FISH nanofishfishdna SPMAFM AFM 2003 SPM pn SPM DNA AFMDNA SPM 2003 1986 AFM 20 SPM 20 SPM SPM SPM SEM
29 SPM SEM SPM SPM SPM NEDO 1 2002. 2 T. Ushiki, S. Yamamoto, J. Hitomi, S. Ogura, T. Umemoto and M. Shigeno: Atomic force microscopy of living cells, Jpn, J. Appl. Phys. 39, 3761-3764 2000. 3 M. Nagayama, H. Haga and K. Kawabata: Drastic change of local stiffness distribution correlating to cell migration in living fibroblasts, Cell Motil Cytoskeleton, 50, 173-179 2001. 4 55 877-879 2002. 5 X-Q. Liu, S. Sugiyama, Q-Y. Xu, T. Kobori, S. Hagiwara and T. Ohtani: Atomic force microscopy study of chromosome surface structure changed by protein extraction Ultramicroscopy 94, 217-223 2003. 6 S. Sugiyama, T. Yoshino, H. Kanahara, M. Shichiri, D. Fukushi and T. Ohtani: Effects of acetic acid treatment on plant chromosome structures analyzed by atomic force microscopy, Anal. Biochem, 324, 39-44 2004.
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31 Resolution using Scanning Near- Field Optical/Atomic Force Microscopy, Arch. Histol. Cytol., 65, 111-120 2002
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51 1 T.Yoshino, S.Isobe, and Takaaki Maekawa, J.American Oil Chemists Society,79 4 345-349 2002 2 R.Paramawati, T.Yoshino and S.Isobe, Food Sci. Technol. Res.,7 3 191-194 2001 3,,, 31,4,225-231 2001 4 T.Yoshino, S.Isobe and T.Maekawa, J. American Oil Chemists Society, 77 7 699-704, 2000 5 45,156 1998 6 S.Isobe et al.,jarq, 31,2,137-146 1997 7 S.Isobe et al.,proceedings of 11th Annual Meeting of BioEnvironmental Polymer Society,p20 2003 8 2003p.40 4115 8 6 9 Q.Wu,H.Sakabe,S.Isobe,Ind.Eng.Chem.Res.,42,6765-6773,2003
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57 2 3-1,4- CA CA -1,4- CD CD Gellman 8 TritonX-100-CD 7 CitSyn 65 40 -CD 3 -CD 17 Cyclodextrin : CD CD6 CD7 CD8 CD CD CD CD9 CD10 CD14 CD26
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 Zn15,000 4
59 S-S 100 1 CitSyn 6 M 1 30 CA CD Tween40 Tween60 0.05 1 CA 0.6 100 CA 1 5
60 CA 100 45CA; 22-45 CA 7 CA; CA 2 B CAB 16 CitSyn CTAB SB3-1490 3 4 mm DTT 6 M16 S-S CAB CTAB SB3-14DL- CA S-S
61 CA 6 1 Lowry 25 30mg/ml 6 M A280 25 1 1 S-S 40mM DTT 2 4 Tween 40, Tween 60
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, 1 142 l of 3 % CA ( 0.6%, 1 (15,000rpm 5 min CTAB, SB3-14 0.05 0.12 0.1 0.15 70 1 CTAB SB3-14 S-S DL-S-S - :GSSG GSH 10 DL- GSH/GSSG DL- 3 3 CA 0.6 1 CA 0.6 CA
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68 Kit 1 Marston,F.A.O. 1986, Biochem.J., 240, 1-12 2 Wetlaufer,D.B.and Xie,T 1995, Prot.Sci., 4, 1535-1543 3 D.Rozema, et al., J. Am. Chem. Soc., 117, 2373 1995 4 M.Yoshimoto. et al., J.Chromatogr.B, 743, 93 2000 5 Y.Akiyoshi, et al., Bioconjugate Chem., 10, 321 1999 6 R.Kuboi. et al., J.Chromatogr.B, 743, 215 2000 7 BioView, 41, 7 2002 8 D.Rozema, & S.H.Gellman., Biochemistry, 35, 15760 1996 9 T.Takaha, et al., J.Biol.Chem., 271, 2902 1996 10 S.Machida, et al., FEBS lett., 486, 131 2000 11 Q.Xie, et al., Protein Expression and Purification, 32, 68-74 2003 12 2003-304624 2003 13 S.Machida, et al., US patent 10-653687 2003
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74 7 8 700 2500 nm 1 2 Brix0.90 0.95 SEP 0.95 1.12 Brix SEP 5 680 1235 nm 1 2 Brix 0.67 0.71 SEP 0.70 0.76 Brix 5 0.97, SEP:0.50 Brix
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85 1 1994 2 G. S. Birth, G. G. Dull, W. T. Renfroe and S. J. Kays: J. Am. Soc. Hort. Sci., 110, 297-303 1985 3 G. G. Dull, G. S. Birth, D. A. Smittle and R. G. Leffler: J. Food. Sci., 54, 393-395 1989 4 S. Kawano, H. Watanabe and M. Iwamoto: Proceedings of the 2 nd International NIRS Conference Ed. M. Iwamoto and S. Kawano, Korin, Tokyo, Japan, pp.343-352 1990 5 S. Kawano, H. Watanabe and M. Iwamoto: J. Jpn. Soc. Hort. Sci., 61, 445-451 1992 6 6
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