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Titolo:
SERUM-DEPENDENT AND CELL CYCLE-DEPENDENT EXPRESSION FROM A CYTOMEGALOVIRUS-BASED MAMMALIAN EXPRESSION VECTOR
Autore:
BRIGHTWELL G; POIRIER V; COLE E; IVINS S; BROWN KW;
Indirizzi:
SCH MED SCI,DEPT PATHOL & MICROBIOL,CLIC RES UNIT,UNIV WALK BRISTOL BS8 1TD AVON ENGLAND SCH MED SCI,DEPT PATHOL & MICROBIOL,CLIC RES UNIT BRISTOL BS8 1TD AVON ENGLAND
Titolo Testata:
Gene
fascicolo: 1, volume: 194, anno: 1997,
pagine: 115 - 123
SICI:
0378-1119(1997)194:1<115:SACCEF>2.0.ZU;2-9
Fonte:
ISI
Lingua:
ENG
Soggetto:
ENHANCER-PROMOTER; WILMS-TUMOR; WT1; GENE; TRANSCRIPTION; STIMULATION;
Keywords:
CYTOMEGALOVIRUS; EXPRESSION VECTOR; CELL CYCLE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
19
Recensione:
Indirizzi per estratti:
Citazione:
G. Brightwell et al., "SERUM-DEPENDENT AND CELL CYCLE-DEPENDENT EXPRESSION FROM A CYTOMEGALOVIRUS-BASED MAMMALIAN EXPRESSION VECTOR", Gene, 194(1), 1997, pp. 115-123

Abstract

Cytomegalovirus-based mammalian expression vectors are widely used todrive the expression of transfected genes in cultured cells. Immunofluorescent staining of the WT1 protein in 3T3 and 293 cell clones, stably transfected with a cyomegalovirus (CMV) expression vector carrying a cDNA coding for the tumour suppressor protein WT1, showed extreme cell to cell variation in the amount of recombinant protein expressed, indicative of cell cycle dependence. This was investigated further by Western blot and FACS analysis which showed that WT1 protein expressionwas highest in S phase and almost absent in G0/G1. Northern blot analysis of cell clones expressing sense or antisense WT1 cDNAs regulated by the CMV promoter/enhancer showed that RNA expression was also cell cycle-dependent. Western blotting of cells expressing a luciferase reporter gene driven by the CMV promoter/enhancer also showed apparent cell cycle-dependent expression. We further demonstrated that the expression of these gene constructs was serum responsive with a 10-fold increase in expression occurring 2 h after the addition of serum. These results show that the CMV promoter/enhancer system varied in its response to serum and the cell cycle state. Therefore, care must be taken when interpreting any phenotypic alterations (or lack of them) produced in cells transfected with CMV-based expression vectors. (C) 1997 Elsevier Science B.V.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 28/11/20 alle ore 21:45:26