Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
A streamlined process to phenotypically profile heterologous cDNAs in parallel using yeast cell-based assays
Autore:
Tugendreich, S; Perkins, E; Couto, J; Barthmaier, P; Sun, DX; Tang, S; Tulac, S; Nguyen, A; Yeh, E; Mays, A; Wallace, E; Lila, T; Shivak, D; Prichard, M; Andrejka, L; Kim, R; Melese, T;
Indirizzi:
Iconix Pharmaceut, Mt View, CA 94043 USA Iconix Pharmaceut Mt View CA USA94043 Pharmaceut, Mt View, CA 94043 USA
Titolo Testata:
GENOME RESEARCH
fascicolo: 11, volume: 11, anno: 2001,
pagine: 1899 - 1912
SICI:
1088-9051(200111)11:11<1899:ASPTPP>2.0.ZU;2-O
Fonte:
ISI
Lingua:
ENG
Soggetto:
GENE-EXPRESSION PATTERNS; SACCHAROMYCES-CEREVISIAE; ESCHERICHIA-COLI; PROTEIN-KINASE; IN-VIVO; MOLECULAR CLASSIFICATION; SHUTTLE VECTORS; DRUG-RESISTANCE; HUMAN CANCER; DNA;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
65
Recensione:
Indirizzi per estratti:
Indirizzo: Melese, T Univ Calif San Francisco, Ctr Comprehens Canc, San Francisco, CA94143 USA Univ Calif San Francisco San Francisco CA USA 94143 A 94143 USA
Citazione:
S. Tugendreich et al., "A streamlined process to phenotypically profile heterologous cDNAs in parallel using yeast cell-based assays", GENOME RES, 11(11), 2001, pp. 1899-1912

Abstract

To meet the demands of developing lead drugs for the profusion of human genes being sequenced as part of the human genome project, we developed a high-throughput assay construction method in yeast. A set of optimized techniques allows us to rapidly transfer large numbers of heterologous cDNAs from nonyeast plasmids into yeast expression vectors. These high- or low-copy yeast expression plasmids are then converted quickly into integration-competent vectors for phenotypic profiling of the heterologous gene products. The process was validated first by testing proteins of diverse function, such as p38, poly(ADP-ribose) polymerase-1, and PI 3-kinase, by making active-site mutations and using existing small molecule inhibitors of these proteins. For less well-characterized genes, a novel random mutagenesis scheme was developed that allows a combination selection/screen for mutations that retain full-length expression and yet reverse a growth phenotype in yeast. A broad range of proteins in different functional classes has been profiled, with an average yield for growth interference phenotypes of similar to 30%. The ease of manipulation of the yeast genome affords us the opportunity to approach drug discovery and exploratory biology on a genomic scale and shortens assay development time significantly.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 03/04/20 alle ore 20:37:51