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1.
¡ºC. M. Niemeyer, C. A. Mirkin, Eds., “Nanobiotechnology, – Concepts, Applications andPerspectives”, Wiley-VCH, Weinheim (2004).
2. T. Ihara, Y. Takeda, A. Jyo, J. Am. Chem. Soc., 123, 1772 (2001).
3. I. Hemmiliä, V. Laitala, J. Fluoresc., 15, 529 (2005).
4. Y. Kitamura, T. Ihara, Y. Tsujimura, M. Tazaki, A. Jyo, Chem. Lett., 34, 1606 (2005).
5. Y. Kitamura, T. Ihara, Y. Tsujimura, Y. Osawa, M. Tazaki, A. Jyo, Anal. Biochem., 100, 1744 (2006).
6. Y. Kitamura, T. Ihara, Y. Tsujimura, Y. Osawa, D. Sasahara, M. Yamamoto, K. Okada, M. Tazaki, A. Jyo,submitted.
7. S. Delaney, J. Yoo, E. D. A. Stemp, J. K. Barton, Proc. Natl. Acad. Sci. USA, 101, 10511 (2004).
8. S. G. Lokhov, A. A. Koshkin, I. V. Kutyavin, M. P. Mityakin, M. A. Podyminogin, A. V. Lebedev,
Russian J. Bioorg. Chem., 21, 169 (1995).
9. Y. Kitamura, T. Ihara, K. Okada, Y. Tsujimura, Y. Shirasaka, M. Tazaki, A. Jyo, Chem. Commun., 4523
(2005).
10. C. M. Dupureur, J. K. Barton, Inorg. Chem., 36, 33 (1997).
11. T. Ihara, Y. Chikaura, S. Tanaka, A. Jyo, Chem. Commun., 2152 (2002).
12. T. Ihara, S. Tanaka, Y. Chikaura, Nucleic Acids Res., 32, e105 (2004).
13. T. Ihara, Y. Mori, T. Imamura, M. Mukae, S. Tanaka, A. Jyo, Anal. Chim. Acta, 578, 11 (2006).
14. S. Ogasawara, K. Fujimoto, Angew. Chem. Int. Ed., 45, 4512 (2006).
15. T. Ihara, T. Fujii, M. Mukae, Y. Kitamura, J. Am. Chem. Soc., 126, 8880 (2004).

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S. L. Schreiber, Science, 287, 1964-1969 (2000).
M. D. Burke and S. L. Schreiber, Angew. Chem. Int. Ed., 43, 46-58 (2004).
  2004, 57(10) 1054-1057.
P. Arya, R. Joseph, Z. Gan, and B. Rakic, Chem. Biol., 12, 163-180 (2005).
D. S. Tan, Nat. Chem. Biol., 1, 74-84 (2005).
D. Lee, J. K. Sello, and S. L. Schreiber, Org. Lett., 2, 709-712 (2000).
O. Kwon, S. B. Park, and S. L. Schreiber, J. Am. Chem. Soc., 124, 13402-13404 (2002).
M. D. Burke, E. M. Berger, and S. L. Schreiber, Science, 302, 613-618 (2003).
H. Oguri, and S. L. Schreiber, Org. Lett., 7, 47-50 (2005).
A. Padwa, and M. D. Weingarten, Chem. Rev., 96, 223-269 (1996).
A. Padwa, and A. T. Price, J. Org. Chem., 63, 556-565 (1998).
A. Dming, and I. Ugi, Angew. Chem. Int. Ed., 39, 3168-3210 (2000).

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Y\WROC[?IIQ5KN7]3KUL@m/&n1. Matsuura T, Yomo T (2006) In vitro evolution of proteins. J Biosci Bioeng 101: 449-456.
2. Matsuura T, Miyai K, Trakulnaleamsai S, Yomo T, Shima Y, et al. (1999) Evolutionary molecularengineering by random elongation mutagenesis. Nat Biotechnol 17: 58-61.
3. Sunami T, Sato K, Matsuura T, Tsukada K, Urabe I, et al. (2006) Femtoliter compartment in liposomes forin vitro selection of proteins. Anal Biochem 357: 128-136.
4. Matsuura T, Yanagida H, Ushioda J, Urabe I, Yomo T (2007) Nascent chain, mRNA, and ribosome
complexes generated by a pure translation system. Biochem Biophys Res Commun 352: 372-377.
5. Yu W, Sato K, Wakabayashi M, Nakaishi T, Ko-Mitamura EP, et al. (2001) Synthesis of functional
protein in liposome. J Biosci Bioeng 92: 590-593.
6. Ishikawa K, Sato K, Shima Y, Urabe I, Yomo T (2004) Expression of a cascading genetic network withinliposomes. FEBS Lett 576: 387-390.
7. Hanes J, Plückthun A (1997) In vitro selection and evolution of functional proteins by using ribosome
display. Proc Natl Acad Sci USA 94: 4937-4942.
8. Matsuura T, Plückthun A (2003) Selection based on the folding properties of proteins with ribosome
display. FEBS Lett 539: 24-28.
9. Koepf EK, Petrassi HM, Ratnaswamy G, Huff ME, Sudol M, et al. (1999) Characterization of thestructure and function of W --> F WW domain variants: identification of a natively unfolded protein thatfolds upon ligand binding. Biochemistry 38: 14338-14351.
10. Tompa P, Szasz C, Buday L (2005) Structural disorder throws new light on moonlighting. Trends
Biochem Sci 30: 484-489.
11. James LC, Roversi P, Tawfik DS (2003) Antibody multispecificity mediated by conformational diversity.
Science 299: 1362-1367.
12. James LC, Tawfik DS (2003) Conformational diversity and protein evolution--a 60-year-old hypothesis
revisited. Trends Biochem Sci 28: 361-368.
13. Shimizu Y, Inoue A, Tomari Y, Suzuki T, Yokogawa T, et al. (2001) Cell-free translation reconstituted
with purified components. Nat Biotechnol 19: 751-755.
14. Riley M, Abe T, Arnaud MB, Berlyn MK, Blattner FR, et al. (2006) Escherichia coli K-12: acooperatively developed annotation snapshot--2005. Nucleic Acids Res 34: 1-9.
15. Butland G, Peregrin-Alvarez JM, Li J, Yang W, Yang X, et al. (2005) Interaction network containing
conserved and essential protein complexes in Escherichia coli. Nature 433: 531-537.

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Binding and enrichment of Escherichia coli spheroplasts expressing inner membrane tethered scFv
antibodies on surface immobilized antigens.
Jung ST, Jeong KJ, Iverson BL, Georgiou G.
Biotechnol. Bioeng., 98, 39-47 (2007).
APEx 2-hybrid, a quantitative protein-protein interaction assay for antibody discovery andengineering.
Jeong KJ, Seo MJ, Iverson BL, Georgiou G.
Proc. Natl. Acad. Sci. U S A., 104, 8247-52 (2007).
Isolation of engineered, full-length antibodies from libraries expressed in Escherichia coli.
Mazor Y, Van Blarcom T, Mabry R, Iverson BL, Georgiou G.
Nat. Biotechnol., 25, 563-5 (2007).
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Biochemistry, 45, 8488-8498 (2006).
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G. Kolesov, Z. Wunderlich, O. N. Laikova, M. S. Gelfand, and L.
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Proc. Natl. Acad. Sci. USA, 104, 13948-13953 (2007).
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Probing Transcription Factor Dynamics at the Single-Molecule Level in a Living Cell
J. Elf, G-W. Li, and X. S. Xie
Science 316, 1191-1194 (2007).
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ÊºÉ´ßÏÐ¯¨9Î$Z»EÀ×¿Ä°   No.25 (2007 October) 29

A Tunable Genetic Switch Based on RNAi and Repressor Proteins for Regulating Gene Expression in
Mammalian Cells
T. L. Deans, C. R. Cantor, and J. J. Collins
Cell 130, 363-372 (2007).
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3}/£\Xvd³¢p¢tet ;hÈ¸ÂØÙµÈ¿Ø`[³iª$²ÚFussenegger £»ÕÙËNucleic Acid Res. 35,
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A
-10

B

Time (min)
0

10

20

30

40

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60

70

80

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C

Time (min)
0

10

20

30

40

50

60

70

80

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  No.25 (2007 October) 31

-0.2

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-0.6

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kcal/mole of injectant

2
-0.8
0

0

0

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Model: OneSites
Chi^2 = 13915.2
N
1.657 ±0.00684
K
4.329E6±2.432E5
H-1.212E4±70.47
S-10.28

-6
-8

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-12

-0.5 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5-4Model: OneSites
Chi^2 = 35839.2
N
1.949 ±0.00745
K
2.45E7 ±2.972E6
H-1.296E4±84.28
S-9.674

-6
-8
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-0.5 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5

Molar Ratio

Model: OneSites
Chi^2 = 18575.9
N
1.873 ±0.00546
K
1.359E8±2.793E7
H-1.031E4±58.95
S
2.633

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Ca/äøöòíöúûDifferential Scanning Calorimetry: DSCüâz»Îµ?7Ô&Ð&JÔt:;PÔCâzÆÎ Ka âh;ÈàZiÜµITC âz»Îµ&78.Ïn;ÈàZiÒÑÀ0)ÅáÎ»à¶Ô<ÐÆÎµmajor histocompatibility complex class II ûMHC IIüÐñðêîÐÔ&ÔÝ¼Óµñðêî&À MHC éùïæÊÔÛÔÔd¥ITÓ5ÁÂ>ÆµMHC !xÏÔÀ,°Ò1&ÒÑÓÕvÓ\Ïµ·Ôñðêîâ MHC Ó&ÅÉËwSÏ DSC n;â»µ%ñ
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2) Saito, K.; Sarai, A.; Oda, M.; Azuma, T.; Kozono, H., J. Biol. Chem., 2003, 278, 14732.
3) Velazquez-Campoy, A.; Freire, E., Nature Protoc., 2006, 1, 186.
4) Oda, M.; Ito, N.; Tsumuraya, T.; Suzuki, K.; Sakakura, M.; Fujii, I., J. Mol. Biol., 2007, 369, 198.
5) Fukada, H.; Takahashi, K., Proteins, 1998, 33, 159.
6) Oda, M.; Furukawa, K.; Ogata, K.; Sarai, A.; Nakamura, H., J. Mol. Biol., 1998, 276, 571.
7) Frederick, K. K.; Marlow, M. S.; Valentine, K. G.; Wand, A. J., Nature, 2007, 448, 325.
8) Spolar, R. S.; Record, M. T., Jr., Science, 1994, 263, 777.
9) Ababou, A.; Ladbury, J. E., J. Mol. Recognit., 2007, 20, 4.
10) Todd, M. J.; Gomez, J., Anal. Biochem., 2001, 296, 179.

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¡§8°«ÓçÆÃÂãËª¹`d½ÈÜéÔ¥¢¤¡lp7w!ª³9s¡° 1) Filomatori, C. V., Lodeiro, M. F., Alvarez, D. E., Samsa, M. M., Pietrasanta, L., and Gamarnik, A. V. A 5'
RNA element promotes dengue virus RNA synthesis on a circular genome. Genes Dev. 20:2238-49(2006).
2) Alvarez, D. E., Lodeiro, M. F., Luduena, S. J., Pietrasanta, L. I., and Gamarnik, A. V. Long-range
RNA-RNA interactions circularize the dengue virus genome. J. Virol. 79:6631-43 (2005).
3) Harada, K., Martin, S. S., and Frankel, A. D. Selection of RNA-binding peptides in vivo. Nature,
380:175-179 (1996).
4) Peled-Zehavi, H., Horiya, S., Das, C., Harada, K., and Frankel, A. D. Selection of RRE RNA binding
peptides using a kanamycin antitermination assay. RNA, 9:252-261 (2003).

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