Combinatorial protein engineering by incremental truncation
نویسندگان
چکیده
منابع مشابه
Combinatorial protein engineering by incremental truncation.
We have developed a combinatorial approach, using incremental truncation libraries of overlapping N- and C-terminal gene fragments, that examines all possible bisection points within a given region of an enzyme that will allow the conversion of a monomeric enzyme into its functional heterodimer. This general method for enzyme bisection will have broad applications in the engineering of new cata...
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Incremental truncation for the creation of hybrid enzymes (ITCHY) is a novel tool for the generation of combinatorial libraries of hybrid proteins independent of DNA sequence homology. We herein report a fundamentally different methodology for creating incremental truncation libraries using nucleotide triphosphate analogs. Central to the method is the polymerase catalyzed, low frequency, random...
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Many proteins can have their peptide backbone cut by proteolytic or genetic means, yet the two fragments can associate to make an active heterodimer. This ‘‘monomer-to-heterodimer conversion’’ is referred to as protein fragment complementation (PFC). Such complementation is the reverse of evolutionary processes in which domains are recruited and fused at the genetic level. Classic examples of p...
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types of combinatorial libraries, such as random shuffling libraries and artificial alternative splicing libraries. The block shuffling and random point mutations can be introduced simultaneously by performing RMPCR under error-prone conditions. Most of the block sequences in the combinatorial libraries created by RM-PCR encode a long open reading frame and are suitable for protein selection ex...
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ژورنال
عنوان ژورنال: Proceedings of the National Academy of Sciences
سال: 1999
ISSN: 0027-8424,1091-6490
DOI: 10.1073/pnas.96.7.3562