DC Field | Value | Language |
---|---|---|
dc.contributor.author | Polizzi, KM | ko |
dc.contributor.author | Spencer, CU | ko |
dc.contributor.author | Dubey, A | ko |
dc.contributor.author | Matsumura, I | ko |
dc.contributor.author | Lee, JayHyung | ko |
dc.contributor.author | Realff, MJ | ko |
dc.contributor.author | Bommarius, AS | ko |
dc.date.accessioned | 2013-03-08T10:35:46Z | - |
dc.date.available | 2013-03-08T10:35:46Z | - |
dc.date.created | 2012-02-06 | - |
dc.date.created | 2012-02-06 | - |
dc.date.issued | 2005-12 | - |
dc.identifier.citation | JOURNAL OF BIOMOLECULAR SCREENING, v.10, no.8, pp.856 - 864 | - |
dc.identifier.issn | 1087-0571 | - |
dc.identifier.uri | http://hdl.handle.net/10203/92848 | - |
dc.description.abstract | Pooling in directed-evolution experiments will greatly increase the throughput of screening systems, but important parameters such as the number of good mutants created and the activity level increase of the good mutants will depend highly on the protein being engineered. The authors developed and validated a Monte Carlo simulation model of pooling that allows the testing of various scenarios in silico before starting experimentation. Using a simplified test system of 2 enzymes, beta-galactosidase (supermutant, or greatly improved enzyme) and beta-glucuronidase (dud, or enzyme with ancestral level of activity), the model accurately predicted the number of supermutants detected in experiments within a factor of 2. Additional simulations using more complex activity distributions show the versatility of the model. Pooling is most suited to cases such as the directed evolution of new function in a protein, where the background level of activity is minimized, making it easier to detect small increases in activity level. Pooling is most successful when a sensitive assay is employed. Using the model will increase the throughput of screening procedures for directed-evolution experiments and thus lead to speedier engineering of proteins. | - |
dc.language | English | - |
dc.publisher | SAGE PUBLICATIONS INC | - |
dc.subject | IN-VITRO EVOLUTION | - |
dc.subject | BETA-GLUCURONIDASE | - |
dc.subject | DIRECTED EVOLUTION | - |
dc.subject | RAPID EVOLUTION | - |
dc.subject | ACTIVE-SITE | - |
dc.subject | GALACTOSIDASE | - |
dc.subject | STRATEGY | - |
dc.subject | DESIGNS | - |
dc.subject | TOOL | - |
dc.title | Simulation modeling of pooling for combinatorial protein engineering | - |
dc.type | Article | - |
dc.identifier.wosid | 000234407000011 | - |
dc.identifier.scopusid | 2-s2.0-29144520637 | - |
dc.type.rims | ART | - |
dc.citation.volume | 10 | - |
dc.citation.issue | 8 | - |
dc.citation.beginningpage | 856 | - |
dc.citation.endingpage | 864 | - |
dc.citation.publicationname | JOURNAL OF BIOMOLECULAR SCREENING | - |
dc.identifier.doi | 10.1177/10871057105280134 | - |
dc.contributor.localauthor | Lee, JayHyung | - |
dc.contributor.nonIdAuthor | Polizzi, KM | - |
dc.contributor.nonIdAuthor | Spencer, CU | - |
dc.contributor.nonIdAuthor | Dubey, A | - |
dc.contributor.nonIdAuthor | Matsumura, I | - |
dc.contributor.nonIdAuthor | Realff, MJ | - |
dc.contributor.nonIdAuthor | Bommarius, AS | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordAuthor | directed evolution | - |
dc.subject.keywordAuthor | high-throughput screening | - |
dc.subject.keywordAuthor | Monte Carlo simulation | - |
dc.subject.keywordAuthor | protein engineering | - |
dc.subject.keywordPlus | IN-VITRO EVOLUTION | - |
dc.subject.keywordPlus | BETA-GLUCURONIDASE | - |
dc.subject.keywordPlus | DIRECTED EVOLUTION | - |
dc.subject.keywordPlus | RAPID EVOLUTION | - |
dc.subject.keywordPlus | ACTIVE-SITE | - |
dc.subject.keywordPlus | GALACTOSIDASE | - |
dc.subject.keywordPlus | STRATEGY | - |
dc.subject.keywordPlus | DESIGNS | - |
dc.subject.keywordPlus | TOOL | - |
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