PROCESS FOR PRODUCING TETRACYCLINE INDUCIBLE GENE EXPRESSING CELL LINE AND CONDITIONAL GENE KNOCKOUT CELL LINE, AND USES THEREOF

摘要

It is intended to provide a method of screening a so-called Tc inducible gene expressing cell line, in which the expression of a gene of interest is regulated depending on the presence/absence of a Tc compound regardless of the type of cell, easily with high efficiency. A gene-of-interest expression vector for transfecting a gene of interest into the genome of a host cell and a TA expression vector that expresses a transactivator that is switched to be bound or not to be bound to a tet operator sequence depending on the presence or absence of tetracycline are prepared. The gene-of-interest expression vector is a vector with a bicistronic regulatory sequence arranged downstream of the tet operator sequence and promoter sequence and between a gene-of-interest coding sequence and a selection marker coding sequence. These vectors are transfected into a host cell, and cell lines in which the selection marker of the gene-of-interest expression vector is expressed are selected. Thus, inducible gene expressing cell lines can be obtained in which the expression of a gene of interest can be controlled with Tc.