A pgk::hprt fusion as a selectable marker for targeting of genes in mouse embryonic stem cells: disruption of the T-cell receptor delta-chain-encoding gene.
We have constructed a hypoxanthine phosphoribosyl transferase-selectable marker (hprt) under the control of the phosphoglycerate kinase (pgk) promoter. This construct permits cell growth in hypoxanthine/aminopterin/thymidine media and confers 6-thioguanine sensitivity upon mouse Hprt- embryonic stem cells, allowing either positive or negative selection in gene-targeting experiments. We have successfully targeted the gene encoding the T-cell receptor delta-chain using the pgk::hprt fusion for counterselection.
van der Lugt
N
N
Division of Molecular Genetics, The Netherlands Cancer Institute, Amsterdam.
Maandag
E R
ER
te Riele
H
H
Laird
P W
PW
Berns
A
A
D000818:Animals; D015139:Blotting, Southern; D005819:Genetic Markers; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D051379:Mice; D010735:Phosphoglycerate Kinase / Q000235:genetics*; D010957:Plasmids; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D013234:Stem Cells
D005819:Genetic Markers; D016692:Receptors, Antigen, T-Cell, gamma-delta; D011993:Recombinant Fusion Proteins; D007041:Hypoxanthine Phosphoribosyltransferase; D010735:Phosphoglycerate Kinase