Allele-specific PCR analysis for detection of the gld Fas-ligand point mutation.
The discovery of a naturally occurring missense point mutation in the gene encoding Fas-ligand (FasL/CD95L) in generalized lymphoproliferative disease (gld) mice has lead to the characterization of FasL as an important mediator of apoptosis. Further analysis of FasL function can be facilitated by crossing the gld mutation onto other mouse-strains, for example those carrying mutations affecting other molecules involved in apoptosis, or disease-prone genetic backgrounds. The success of this is dependent on a quick and reliable screening method. Here we report an allele-specific PCR for detection of the gld mutation. This approach permits the screening of back-crossed F1 progeny within one day, using whole blood samples as a source of genomic DNA. The technique is fast, robust, easily learnt, and unambiguous.
Hoek
R M
RM
Centenary Institute of Cancer Medicine and Cell Biology, Royal Prince Alfred Hospital, Sydney, NSW, Australia.
Kortekaas
M C
MC
Sedgwick
J D
JD
D000483:Alleles; D000818:Animals; D004252:DNA Mutational Analysis / Q000379:methods; D053222:Fas Ligand Protein; D008024:Ligands; D008232:Lymphoproliferative Disorders / Q000235:genetics; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000235:genetics; D051379:Mice; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D019014:fas Receptor / Q000032:analysis* / Q000235:genetics
D053222:Fas Ligand Protein; C505658:Fasl protein, mouse; D008024:Ligands; D008562:Membrane Glycoproteins; D019014:fas Receptor