Combinatorial cassette mutagenesis as a probe of the informational content of protein sequences

John F. Reidhaar-Olson, Robert T. Sauer

Research output: Contribution to journalArticle

271 Citations (Scopus)

Abstract

A method of combinatorial cassette mutagenesis was designed to readily determine the informational content of individual residues in protein sequences. The technique consists of simultaneously randomizing two or three positions by oligonucleotide cassette mutagenesis, selecting for functional protein, and then sequencing to determine the spectrum of allowable substitutions at each position. Repeated application of this method to the dimer interface of the DNA-binding domain of λ repressor reveals that the number and type of substitutions allowed at each position are extremely variable. At some positions only one or two residues are functionally acceptable; at other positions a wide range of residues and residue types are tolerated. The number of substitutions allowed at each position roughly correlates with the solvent accessibility of the wild-type side chain.

Original languageEnglish (US)
Pages (from-to)53-57
Number of pages5
JournalScience
Volume241
Issue number4861
StatePublished - 1988
Externally publishedYes

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Insertional Mutagenesis
Protein Sequence Analysis
Oligonucleotides
Proteins
DNA

ASJC Scopus subject areas

  • General

Cite this

Reidhaar-Olson, J. F., & Sauer, R. T. (1988). Combinatorial cassette mutagenesis as a probe of the informational content of protein sequences. Science, 241(4861), 53-57.

Combinatorial cassette mutagenesis as a probe of the informational content of protein sequences. / Reidhaar-Olson, John F.; Sauer, Robert T.

In: Science, Vol. 241, No. 4861, 1988, p. 53-57.

Research output: Contribution to journalArticle

Reidhaar-Olson, JF & Sauer, RT 1988, 'Combinatorial cassette mutagenesis as a probe of the informational content of protein sequences', Science, vol. 241, no. 4861, pp. 53-57.
Reidhaar-Olson, John F. ; Sauer, Robert T. / Combinatorial cassette mutagenesis as a probe of the informational content of protein sequences. In: Science. 1988 ; Vol. 241, No. 4861. pp. 53-57.
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