A new retroviral vector, CA1, to identify and select for cells expressing an inserted gene in vitro and in vivo.
Gene
; 196(1-2): 187-9, 1997 Sep 01.
Article
em En
| MEDLINE
| ID: mdl-9322757
ABSTRACT
A new retroviral vector has been constructed that expresses genes encoding three different activities from a single transcript. This feature has been exploited to enable the efficient marking and selection of cells that express a gene of interest. The marker gene lacZ, encoding beta-galactosidase, and neo, encoding neomycin phosphotransferase, for selection by the antibiotic G418, are expressed as a fusion, beta Geo. The expression of beta Geo is coordinated with expression of a gene of interest at the mRNA level using an Internal Ribosome Entry Site (IRES) from the Encephalomyocarditis Virus (EMCV). The IRES promotes cap-independent initiation of translation therefore two reading frames can be translated from a single transcript. In vitro, the vector has been shown to confer beta-galactosidase activity, transformation by v-src and resistance to G418, following infection of cells. To show that the retrovirus was able to mark infected cells in vivo, cells infected with the retrovirus were transplanted into mouse mammary gland where they grew and were successfully located by staining for beta-galactosidase over 2 months after transplantation.
Buscar no Google
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Retroviridae
/
Seleção Genética
/
Proteínas Recombinantes
/
Vetores Genéticos
Limite:
Animals
Idioma:
En
Revista:
Gene
Ano de publicação:
1997
Tipo de documento:
Article
País de afiliação:
Reino Unido