Abstract
In the early 1970s, recombinant DNA technology made in vitro manipulation of genetic information possible. In the early 198Os, advances in solid phase synthetic chemistry made the production of oligonucleotides of defined sequence rapid and economical. Since then, methods of site-directed mutagenesis (SDM) built on these advances have been refined to the point that unlimited variations of a gene sequence can be created given sufficient time. Furthermore, commercially available kits have opened the realm of site-directed mutagenesis to laboratories that are not specialists in DNA molecular biology. The ability to introduce specific mutations into a gene and then express and study the altered protein has provided a powerful experimental tool for studying the relationship between amino acid sequence and protein structure and stability.
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References
Brandts, J. F. (1964) J. Am. Chem. Sot. 86, 4302–4314.
Kauzmann, W. (1959) Adv. Protein Chem. 14, 1–63.
Tanford, C. (1962) J. Am. Chem. Sot. 84, 4240–4247.
Tanford, C. (1970) Adv. Protein Chem. 24, 1–95.
Pantoliano, M. W., Whitlow, M., Wood, J. F., Dodd, S. W., Hardman, K. D., Rollence, M. L., and Bryan, P. N. (1989) Biochemistry 28, 7205–7213.
Leatherbarrow, R. J. and Fersht, A. R. (1986) Protein Eng. 1, 7–16.
Matthews, B. W. (1987) Biochemistry 26, 6885–6888.
Goldenberg, D. P. (1988) Ann. Rev. Biophys. Biophys. Chem. 17, 481–507.
Alber, T. (1989) Ann. Rev. Biochem. 58, 765–798.
Wells, J. A., Ferrari, E., Henner, D. J., Estell, D. A., and Chen, E. Y. (1983) Nucleic Acids Res. 11, 7911–7925.
Vasantha, N., Thompson, L. D., Rhodes, C., Banner, C., Nagle, J., and Filpula, D. (1984) J. Bucteriol. 159, 811–819.
Fahnestock, S. R. and Fisher, K. E. (1987) Appl. Environ. Microbial. 53, 379–384.
Bryan, P., Alexander, P., Strausberg, S., Schwarz, F., Wang, L., Gilliland, G., and Gallagher, D. T. (1992) Biochemistry 31, 4937–4945.
DelMar, E., Largman, C., Brodnck, J., and Geokas, M (1979) Anal. Biochem. 99, 316–320.
Wiseman, T., Williston, S., Brandts, J. F., and Lin, L.-N. (1989) Anal. Biochem. 179, 131–137.
Gallagher, T. D., Bryan, P., and Gilliland, G. (1993) Proteins: Str. Funct. Gen. 16, 205–213.
Matthews, B. W., Weaver, L. H., and Kester, W. R. (1974) J. Biol. Chem. 249, 8030–8044.
Betzel, C., Teplyakov, A. V., Harutyunyan, E. H., Sanger, W., and Wilson, K. S. (1990) Protein Eng. 3, 161–172.
Gros, P., Kalk, K. H., and Hol, W. G. J. (1991) J. Biol. Chem. 266, 2953–2961.
Ikemura, H., Takagi, H., and Inouye, M. (1987) J. Biol. Chem. 262, 7859–7864.
Finzel, B. C., Howard, A. J., and Pantoliano, M. W. (1986) J. Cell. Biochem. Suppl. 10A, 272.
Pantoliano, M. W., Whitlow, M., Wood, J. P., Rollence, M. L., Finzel, B. C., Gilliland, G., Poulos, T. L., and Bryan, P. N. (1988) Biochemistry 27, 831l–8317.
Bryan, P. N., Rollence, M. L., Pantoliano, M. W., Wood, J., Finzel, B. C., Gilliland, G. L., Howard, A. J., and Poulos, T. L. (1986) Prot. Str. Funct. Gen. 1, 326–334.
Jones, T. A. (1978) J. Appl. Crystallogr. 11, 268–272.
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Bryan, P.N. (1995). Site-Directed Mutagenesis to Study Protein Folding and Stability. In: Shirley, B.A. (eds) Protein Stability and Folding. Methods in Molecular Biology™, vol 40. Humana Press. https://doi.org/10.1385/0-89603-301-5:271
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DOI: https://doi.org/10.1385/0-89603-301-5:271
Publisher Name: Humana Press
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