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Proteins to Order Use of Synthetic DNA to Generate Site-Specific Mutations

Cohen, L. and Molin, C. and Itakura, K. and Riggs, A. D. and McFarland, G. Dalbadie and Richards, J. H. (1982) Proteins to Order Use of Synthetic DNA to Generate Site-Specific Mutations. In: IUPAC--International Union of Pure and Applied Chemistry 28th Macromolecular Symposium. International Union of Pure and Applied Chemistry , p. 335. http://resolver.caltech.edu/CaltechAUTHORS:20150506-120534864

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Abstract

The ability to cause specific changes in the amino acid sequences of proteins would greatly advance studies on the influence of protein structure on biochemical function. If the desired changes can once be made in the nucleic acid which encodes the protein, one can use cloning in an appropriate microorganism to produce essentially limitless quantities of the mutant protein. We describe here the application of oligonucleotide-directed site-specific mutagenesis to accomplish this objective for the enzyme B-lactamase, the gene for which is contained in the plasmid pBR322. The method uses a procedure to screen for mutant clones which depends on the DNA in the various colonies and not on the properties of the mutant protein; the method can, therefore, be widely applied and does not require, in each separate case, the development of a screening procedure which depends on some phenotypic difference between mutant and wild-type protein.


Item Type:Book Section
Additional Information:© 1982 International Union of Pure and Applied Chemistry.
Record Number:CaltechAUTHORS:20150506-120534864
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20150506-120534864
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:57277
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:06 May 2015 22:51
Last Modified:06 May 2015 22:51

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