Abstract
A new approach for the synthesis of randomized DNA sequences containing the 20 codons corresponding to all natural amino acids is described. The strategy is based on the use of dinucleotide phosphoramidite building blocks within a resin-splitting procedure. Through this protocol, a minimal number of seven dimers is sufficient to encode all 20 natural amino acids. This synthesis procedure is extremely flexible and allows codon usage from different hosts to be accommodated.
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Selected References
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- Braunagel M., Little M. Construction of a semisynthetic antibody library using trinucleotide oligos. Nucleic Acids Res. 1997 Nov 15;25(22):4690–4691. doi: 10.1093/nar/25.22.4690. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cormack B. P., Struhl K. Regional codon randomization: defining a TATA-binding protein surface required for RNA polymerase III transcription. Science. 1993 Oct 8;262(5131):244–248. doi: 10.1126/science.8211143. [DOI] [PubMed] [Google Scholar]
- Glaser S. M., Yelton D. E., Huse W. D. Antibody engineering by codon-based mutagenesis in a filamentous phage vector system. J Immunol. 1992 Dec 15;149(12):3903–3913. [PubMed] [Google Scholar]
- Huang W., Santi D. V. Identification of biologically active mutants by combinatorial cassette mutagenesis: exclusion of wild-type codon from degenerate codons. Anal Biochem. 1994 May 1;218(2):454–457. doi: 10.1006/abio.1994.1206. [DOI] [PubMed] [Google Scholar]
- Kayushin A. L., Korosteleva M. D., Miroshnikov A. I., Kosch W., Zubov D., Piel N. A convenient approach to the synthesis of trinucleotide phosphoramidites--synthons for the generation of oligonucleotide/peptide libraries. Nucleic Acids Res. 1996 Oct 1;24(19):3748–3755. doi: 10.1093/nar/24.19.3748. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lam K. S., Salmon S. E., Hersh E. M., Hruby V. J., Kazmierski W. M., Knapp R. J. A new type of synthetic peptide library for identifying ligand-binding activity. Nature. 1991 Nov 7;354(6348):82–84. doi: 10.1038/354082a0. [DOI] [PubMed] [Google Scholar]
- Lyttle M. H., Napolitano E. W., Calio B. L., Kauvar L. M. Mutagenesis using trinucleotide beta-cyanoethyl phosphoramidites. Biotechniques. 1995 Aug;19(2):274–281. [PubMed] [Google Scholar]
- Ono A., Matsuda A., Zhao J., Santi D. V. The synthesis of blocked triplet-phosphoramidites and their use in mutagenesis. Nucleic Acids Res. 1995 Nov 25;23(22):4677–4682. doi: 10.1093/nar/23.22.4677. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Short J. M., Fernandez J. M., Sorge J. A., Huse W. D. Lambda ZAP: a bacteriophage lambda expression vector with in vivo excision properties. Nucleic Acids Res. 1988 Aug 11;16(15):7583–7600. doi: 10.1093/nar/16.15.7583. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sondek J., Shortle D. A general strategy for random insertion and substitution mutagenesis: substoichiometric coupling of trinucleotide phosphoramidites. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3581–3585. doi: 10.1073/pnas.89.8.3581. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Virnekäs B., Ge L., Plückthun A., Schneider K. C., Wellnhofer G., Moroney S. E. Trinucleotide phosphoramidites: ideal reagents for the synthesis of mixed oligonucleotides for random mutagenesis. Nucleic Acids Res. 1994 Dec 25;22(25):5600–5607. doi: 10.1093/nar/22.25.5600. [DOI] [PMC free article] [PubMed] [Google Scholar]