The donor splice site mutation in NFkappaB-inducing kinase of alymphoplasia (aly/aly) mice.

Abstract

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.