The donor splice site mutation in NFkappaB-inducing kinase of alymphoplasia (aly/aly) mice.
The alymphoplasia (aly/aly) mouse has a spontaneous mutation maintained on a C57BL/6xAEJ ( H-2(b)) background that results in an absence of extrasplenic secondary lymphoid tissues. The cDNA defect has previously been shown to reside in a point mutation causing a G855R substitution in NFkappaB-inducing kinase (NIK). Since the aly/aly female cannot lactate, the strain must be bred by intercrossing heterozygous females with homozygous males and the offspring typed by serum IgA levels at the age of 4-6 weeks. We originally determined the genomic location of the alymphoplasia mutation by sequencing boundaries of regions homologous to human NIK exons, although recently the entire genomic sequence of murine C57BL/6 NIK has become available through the mouse genome project. The aly mutation is at position -1 of an intron donor consensus splice site. Exon-connexion PCR confirmed that splicing does occur across this site. Using the genomic information, we also developed a method of PCR typing of aly/aly mice from tail clips, and used this to derive an aly/aly muMT double-mutant strain in which antibody independent typing is essential. Genetic typing should considerably simplify husbandry and manipulation of the aly/aly genetic background, which is widely used as a recipient in lymphocyte transfer experiments to permit examination of the relative role of secondary lymphoid structures in immune responses.
Macpherson
Andrew J
AJ
Institute of Experimental ImmunologyUniversitätsspital, Schmelzbergstrasse 12, 8091 Zürich Switzerland. amacpher@pathol.unizh.ch
Uhr
Therese
T
D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000737:chemistry; D005260:Female; D008198:Lymph Nodes / Q000002:abnormalities*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010581:Peyer's Patches / Q000002:abnormalities*; D017354:Point Mutation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000097782:NF-kappaB-Inducing Kinase
D018076:DNA, Complementary; D017346:Protein Serine-Threonine Kinases; D012333:RNA, Messenger; D000097782:NF-kappaB-Inducing Kinase