8

8. El diseño 'irracional'. Problemas del diseño racional. Bibliotecas proteicas. Identificación y selección. Estrategias de selección.

Bibliografía:

Metodología

3D structural information as a guide to protein engineering using genetic selection. P. Kast & D. Hilvert. COSB 7, 470-479 (1997).

Combinatorial protein design: strategies for screening protein libraries. H. Zhao & F.H. Arnold. COSB 7, 480-485 (1997).

Combinatorial chemistry. JN Abelson in Methods in Enzymology 267 (1996)

DNA shuffling by random fragmentation and reassembly: in vitro recombination for molecular evolution. WPC Stemmer. Bio-technology (1995) 13: 549-553.

Phage display: protein engineering by directed evolution. KT O'Neil, RH Hoess. COSB 5, 443-449 (1995)

Design and use of a phage display library. A. Pini, F. Viti, A. Santucci, B. CArnemolla, L. Zardi, P. Neri, D, Neri. (1998). J. Biol. Chem. 273, 21769-21776

In vitro selection and evolution of functional proteins by using ribosome display. J Hanes & A Plückthun. PNAS(1997) 94: 4937-4942

Ejemplos

Novel human DNA alkyltransferases obtained by random substitution and genetic selection in bacteria. FC Christians, LA Loeb. PNAS (1996) 93: 6124-6128.

Improved green fluorescence protein by molecular evolution using DNA shuffling. A Cremeri, EA Whitehorn, E Tale, WPC Stemmer. Nat. Biotechnol. (1996) 14: 315-319.

Guided evolution of enzymes with new substrate specificity. AS Hawrani, RB Sessions, KM Moreton, JJ Holbrook. (1996) J. Mol. Biol. 264: 97-110.

Directed evolution of para-nitrobenzyl esterase for aqueous-organic solvents. JC Moore, FH Arnold, Nat. Biotech. 14:458-467 (1996)

Selection of RNA-binding peptides in vivo. K. Harada, SS Martin, AD Frankel. Nature 380: 175-179 (1996)

In vitro evolution of thermodinamically stable turns. HX Zhou, RH Hoess, WF DeGrado. Nat. Struct. Biol. 3, 446-451 (1996)

A general strategy for selecting high-affinity zinc finger proteins for diverse DNA target sites. H.A. Greisman, C.O. Pabo. (1997). Science 275, 657-661.

A strategy for the isolation of catalytic activities from repertoires of enzymes diaplayed on phage. S. Demartis, A. Huber, F. Viti, L. Lozzi, L. Giovannoni, P. Neri, G. Winter, D. Neri. (1999). J. Mol. Biol. 286, 617-633.