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Titolo:
Exogenous peptide and protein expression levels using retroviral vectors in human cells
Autore:
Sandrock, TM; Risley, B; Richards, BT; Poritz, MA; Austin, HA; Yoo, S; Kim, MKH; Roth, B; Repetny, K; Hsu, F; Stump, M; Teng, DHF; Kamb, A;
Indirizzi:
Deltagen Proteom, Salt Lake City, UT 84108 USA Deltagen Proteom Salt LakeCity UT USA 84108 Salt Lake City, UT 84108 USA
Titolo Testata:
MOLECULAR THERAPY
fascicolo: 5, volume: 4, anno: 2001,
pagine: 398 - 406
SICI:
1525-0016(200111)4:5<398:EPAPEL>2.0.ZU;2-G
Fonte:
ISI
Lingua:
ENG
Soggetto:
GREEN FLUORESCENT PROTEIN; POSTTRANSCRIPTIONAL REGULATORY ELEMENT; GENETIC SUPPRESSOR ELEMENTS; HUMAN PAPILLOMAVIRUS TYPE-16; MAMMALIAN-CELLS; HALF-LIFE; RESISTANCE; IMMORTALIZATION; TRANSGENES; PRODUCT;
Keywords:
transdominant; peptide; concentration; E7; retroviral; retinoblastoma; cell line;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
34
Recensione:
Indirizzi per estratti:
Indirizzo: Sandrock, TM Deltagen Proteom, 615 Arapeen Dr,Suite 300, Salt Lake City, UT 84108 USA Deltagen Proteom 615 Arapeen Dr,Suite 300 Salt Lake City UT USA84108
Citazione:
T.M. Sandrock et al., "Exogenous peptide and protein expression levels using retroviral vectors in human cells", MOL THER, 4(5), 2001, pp. 398-406

Abstract

Pseudotyped retroviral vectors combine the advantages of broad host range,high expression, stable chromosomal integration, and ease of preparation. These vectors greatly facilitate delivery into mammalian cells of sequencesencoding individual peptide inhibitors-including those with therapeutic utility-and inhibitor libraries. However, retroviral vectors vary in behavior, particularly with respect to expression levels in different cell lines. Expression level is especially important in transdominant experiments because the concentration of an inhibitor (for example, an expressed peptide) is one of the key determinants in the degree of complex formation between the inhibitor and its target. Thus, inhibitor concentration should have an impact on the expressivity and/or penetrance of an induced phenotype. Here, we compare several retroviral vectors and human cell lines for relative expression levels using a green fluorescent protein reporter. We show for a subset of these lines that cellular protein concentrations produced by single-copy vectors range up to about 2 muM. We also examine other variables that contribute to expression level, such as the nature of the expressed protein'scarboxy terminus. Finally, we test the effect of increased concentration on phenotype with a nine-amino-acid peptide derived from the human papillomavirus protein E7 which overcomes E7-mediated cell growth.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 29/03/20 alle ore 16:13:41