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Titolo:
Generation of RCAS vectors useful for functional genomic analyses
Autore:
Loftus, SK; Larson, DM; Watkins-Chow, D; Church, DM; Pavan, WJ;
Indirizzi:
NHGRI, Mouse Embryol Sect, Genet Dis Res Inst, NIH, Bethesda, MD 20892 USANHGRI Bethesda MD USA 20892 net Dis Res Inst, NIH, Bethesda, MD 20892 USA NIH, Natl Ctr Biotechnol Informat, Bethesda, MD 20892 USA NIH Bethesda MDUSA 20892 Ctr Biotechnol Informat, Bethesda, MD 20892 USA
Titolo Testata:
DNA RESEARCH
fascicolo: 5, volume: 8, anno: 2001,
pagine: 221 - 226
SICI:
1340-2838(20011031)8:5<221:GORVUF>2.0.ZU;2-4
Fonte:
ISI
Lingua:
ENG
Soggetto:
ROUS-SARCOMA VIRUS; SITE-SPECIFIC RECOMBINATION; RETROVIRAL VECTORS; AVIAN-SARCOMA; GENE-TRANSFER; DNA-SYNTHESIS; CELL-LINE; SYSTEM; MICE; INTEGRATION;
Keywords:
RCAS; tv-a; retrovirus; recombination; Gateway; gene expression;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
23
Recensione:
Indirizzi per estratti:
Indirizzo: Loftus, SK NHGRI, Mouse Embryol Sect, Genet Dis Res Inst, NIH, Bldg 49,Room 4A67,49 Convent Dr,MSC4472, Bethesda, MD 20892 USA NHGRI Bldg 49,Room 4A67,49 Convent Dr,MSC4472 Bethesda MD USA 20892
Citazione:
S.K. Loftus et al., "Generation of RCAS vectors useful for functional genomic analyses", DNA RES, 8(5), 2001, pp. 221-226

Abstract

Avian leukosis type A virus-derived retroviral vectors have been used to introduce genes into cells expressing the corresponding avian receptor tv-a. This includes the use of Replication-Competent Avian sarcoma-leukosis virus (ASLV) long terminal repeat (LTR) with Splice acceptor (RCAS) vectors in the analysis of avian development, human and murine cell cultures, murine cell lineage studies and cancer biology. Previously, cloning of genes into this virus was difficult due to the large size of the vector and sparse cloning sites. To overcome some of the disadvantages of traditional cloning using the RCASBP-Y vector, we have modified the RCASBP-Y to incorporate "Gateway" site-specific recombination cloning of genes into the construct, eitherwith or without HA epitope tags. We have found the repetitive "att" sequences, which are the targets for site-specific recombination, do not impair the production of infectious viral particles or the expression of the gene of interest. This is the first instance of site-specific recombination beingused to generate retroviral gene constructs. These viral constructs will allow for the efficient transfer and expression of cDNAs needed for functional genomic analyses.

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Documento generato il 05/04/20 alle ore 13:39:48