Glucocorticoid-inducible expression of a glutamine synthetase-CAT-encoding fusion plasmid after transfection of intact chicken retinal explant cultures

• Published in: Gene Vol. 89; no. 2; pp. 259 - 263
• Main Authors: Pu, Haifeng, Young, Anthony P.
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 14.05.1990; Amsterdam Elsevier; New York, NY
• Subjects: Animals; Biological and medical sciences; Biotechnology; Chick Embryo; Chloramphenicol O-Acetyltransferase - genetics; Chloramphenicol O-Acetyltransferase - metabolism; Dexamethasone - pharmacology; Electric Stimulation; electroporation; Fundamental and applied biological sciences. Psychology; Gene Expression - drug effects; gene transfer of intact tissue; Genetic engineering; Genetic technics; Glutamate-Ammonia Ligase - genetics; Glutamate-Ammonia Ligase - metabolism; Methods. Procedures. Technologies; Plasmids; Promoter Regions, Genetic; Recombinant DNA; Recombinant Fusion Proteins - metabolism; Retina - drug effects; Retina - metabolism; Transcription, Genetic - drug effects; Transfection; PBS; nt; gene transfer of intact tissue; Cm; Recombinant DNA; Glns; LTR; bp; βGal; E8 or E10; tsp; DMEM; RSV; Dex; CAT; FCS; kb; tk; electroporation; GRE; Heterologous system; Enzyme; Steroid hormone; Induction; Tissue culture; Electroporation; Retina; Glutamine synthetase; Gene expression; Glucocorticoid; Genetic transfer; Vertebrata; Transfection; Genetic engineering; Chicken; Aves; Hybrid gene
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/0378-1119(90)90014-I

In this communication we demonstrate that gene transfer methodology can be applied to study gene expression in intact retinal explant cultures. The appropriate enzyme activity is observed in extracts obtained after electroporation of embryonic day-10 chicken retina with plasmids containing the chloramphenicol acetyltransferase-encoding or γ-galactosidase-encoding reporter genes under transcriptional control by the Rous sarcoma virus long terminal repeat. Similar results are obtained using Ca · phosphate-mediated gene transfer. Moreover, it has been previously established that glucocorticoid hormones stimulate transcription of glutamine synthetase (Glns) mRNA in embryonic retina. We report here that, based on the results of gene transfer experiments with chimeric plasmids containing 5′ - flanking DNA derived from the cloned chicken Glns- encoding gene ( Glns), essential glucocorticoid response elements residebetween approx. 1.3 kb and 2.5 kb upstream from the Glns transcription start point. These data show that transfection of explant cultures can provide a useful approach to the study of gene expression in complex systems.