Improved expression vector for consistent cellular expression of the tet on repressor in multiple cell types

摘要

A vector having the Protein Translation Peptide Elongation Factor-1alpha (EF-1alpha) promoter and nucleic acids encoding a reverse tetracycline-controlled activator is provided, wherein the expression of the activator is under the control of the EF-1alpha promoter. In addition, a method of generating a reverse tetracycline-controlled transactivator expression system for inducible tetracycline-regulated gene expression is provided that consists of: (a) isolation of a DNA fragment encoding the reverse tetracycline-controlled transactivator, (b) isolation of a DNA fragment containing the EF-1alpha promoter, (c) subcloning of these DNA fragments into a plasmid or other suitable vector to create a vector in which the reverse tetracycline-controlled transactivator is operably linked to the EF-1alpha promoter.