Production of Cells with Targeted Integration of Gene Variants of Human ABC Transporter for Stable and Regulated Expression Using the Flp Recombinase System - Protein Misfolding and Cellular Stress in Disease and Aging - page 137

(ATCC) cell lines, for example, HEK293, CV-1,
CHO-K1, BHK, NIH/3T3, or Jurkat, to stably express
the lac Z-Zeocin fusion gene. Each cell line contains a
single integrated F lp R ecombination T Target (FRT) site.
The FRT site, originally isolated from Saccharomyces cer-
evisiae , serves as a binding site for Flp recombinase and
has been well characterized ( 29, 35-37 ). The minimal
FRT site consists of a 34-bp sequence containing two
13-bp imperfect inverted repeats separated by an 8-bp
spacer that includes an Xba I restriction site (Fig. 2a ). An
Fig. 2. DNA sequence of FRT site ( a ), schematic illustration of the stable expression of ABCG2 in Flp-In-293 cells ( b ), FISH
mapping ( c ), and multicolor-FISH analysis ( d ). Flp-In-293 cells were co-transfected with the pcDNA5/FRT vector carrying