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Titolo:
EVALUATION OF THE CONTINGENT REPLICATION ASSAY (CRA) AND ITS APPLICATION TO THE STUDY OF THE GENERAL TRANSCRIPTION INITIATION-FACTOR, TFIIF
Autore:
NALLUR GN; VASAVADA HA; SANKHAVARAM PR; XU WJ; WEISSMAN SM;
Indirizzi:
YALE UNIV,SCH MED,BOYER CTR MOLEC MED,DEPT GENET,295 CONGRESS AVE NEWHAVEN CT 06356 YALE UNIV,SCH MED,BOYER CTR MOLEC MED,DEPT GENET,295 CONGRESS AVE NEWHAVEN CT 06356
Titolo Testata:
Nucleic acids research
fascicolo: 16, volume: 21, anno: 1993,
pagine: 3867 - 3873
SICI:
0305-1048(1993)21:16<3867:EOTCRA>2.0.ZU;2-R
Fonte:
ISI
Lingua:
ENG
Soggetto:
PROTEIN PROTEIN INTERACTIONS; RNA POLYMERASE-II; MAMMALIAN-CELLS; DNA; CDNA; CLONING; PLASMID; SYSTEM; VECTOR; FAMILY;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
28
Recensione:
Indirizzi per estratti:
Citazione:
G.N. Nallur et al., "EVALUATION OF THE CONTINGENT REPLICATION ASSAY (CRA) AND ITS APPLICATION TO THE STUDY OF THE GENERAL TRANSCRIPTION INITIATION-FACTOR, TFIIF", Nucleic acids research, 21(16), 1993, pp. 3867-3873

Abstract

The Contingent replication assay (CRA) is a rapid assay for the screening and isolation of cDNAs by protein-protein or protein-DNA interactions in mammalian cells. The method has been shown to enrich a plasmidcontaining a cDNA encoding the bacterial replication-related protein,R6K, from a mixture of two plasmids. In this report we present data illustrating the sensitivity and selectivity of the method. Using the small subunit of TFIIF (Rap30) as a target, we demonstrate the enrichment of a clone encoding the large subunit, Rap74, from a cDNA library. Additional cDNA clones including human Rap30 and an anonymous cDNA clone homologous to members of the human cdc2 kinase family were enrichedand isolated by a modified screening approach. The structure of theseadditional clones suggest that the CRA enriches for products that interact not only directly with the target protein but also through bridging by endogenous proteins.

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Documento generato il 05/08/20 alle ore 02:11:42