Construction and hormone regulation of a novel retroviral vector.
We report the analysis of a self-inactivating retroviral vector, constructed to allow inducible gene expression of inserted sequences from the mouse mammary tumour virus hormonal response element. The cloning strategy has been designed to allow for ease of insertion of the genes of interest. The vector contains the aph gene, allowing geneticin-resistance selection in mammalian cells. We have characterised dexamethasone (Dex)-induced increase in gene expression using the reporter gene encoding chloramphenicol acetyltransferase (CAT) inserted into the retroviral vector. We observe low basal levels of CAT activity in infected cells which is increased up to 50-fold by induction with Dex. The induction of pooled clones is 13.3-fold. Variation in Dex-induced CAT activity is observed in independently infected clones, which is not explained by proviral copy number.
Mee
P J
PJ
CRC Department of Medical Oncology, Glasgow University, U.K.
Brown
R
R
D000818:Animals; D015500:Chloramphenicol O-Acetyltransferase; D003001:Cloning, Molecular; D003907:Dexamethasone / Q000494:pharmacology; D004352:Drug Resistance, Microbial / Q000235:genetics; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors; D005839:Gentamicins; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics*; D051379:Mice; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000187:drug effects; D012091:Repetitive Sequences, Nucleic Acid; D012190:Retroviridae / Q000235:genetics*
D005839:Gentamicins; D003907:Dexamethasone; C010680:antibiotic G 418; D015500:Chloramphenicol O-Acetyltransferase