The donor splice site mutation in NFκB-inducing kinase of alymphoplasia (aly/aly) mice
2002
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 NFκB-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 μMT 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.
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