Introduction of the glucocorticoid binding sequences into the expression vector p delta SVE-CAT and its effect on the CAT gene expression in mammalian cells.
We have demonstrated that synthetic oligonucleotide representing glucocorticoid responsive element (GRE I) of MMTV inserted into the enhancerless early promoter of SV40 in p delta SVE-CAT expression vector, enhances transient expression of chloramphenicol acetyltransferase gene in HeLa and hepatoma cells cultivated in the presence of dexamethasone. The following changes in the structure of the core sequences (GTTACAAACTGTTCT) of the synthesized GRE eliminated its enhancing ability: i, changes in the left end of the core sequences from GTTACAAAATGTTCT to TCTTCAAACTGTTCT or to TACTCAAACTGTTCT; ii, the increase of gap between TGTTCT and the inverted repeat of this sequence. The above changes did not eliminate specific binding of glucocorticoid receptor to the synthetic oligonucleotides studied.
Sokół-Misiak
W
W
Department of Molecular Biology, Polish Academy of Sciences, Mroków.
Markiewicz
W T
WT
Szydłowska
B
B
Petrenko
A
A
D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics*; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D005938:Glucocorticoids / Q000737:chemistry / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008114:Liver Neoplasms, Experimental; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D051381:Rats; D014158:Transcription, Genetic
D005938:Glucocorticoids; D015500:Chloramphenicol O-Acetyltransferase