Title: Identification through ENU genome-wide mutagenesis of a novel mutant mouse that expresses a non-functional induced T cell kinase (Itk) with 8-amino acid insertion (33.30)
Abstract: Abstract With the human genome sequenced, the race is on to understand the functions of the 30,000 or so genes. Ethyl-nitrosourea (ENU) was used to treat C57BL/6 (B6) male mice, followed by a 3-generation breeding scheme to generate G3 mice that were screened for immunological phenotypes under recessive genetic control. The primary phenotypes of one mutant, P-135, were increased CD44hi representation among both CD4 and CD8 T cells and highly elevated serum IgE. Recessive mode of inheritance was confirmed by mating an affected G3 male to wildtype B6 females to generate F2 offspring. Affected G3 mice were outcrossed to generated F2 offspring and the affected F2 mice were subjected to linkage analysis by haplotype interval mapping (Neuhaus and Beier, 1998. Mammalian Genome 9, 150-154). The mutant gene was found to be linked to chromosome 11. Through DNA sequencing of genes located within the linked region, a point mutation (T to C) in the 12th intron was identified. This point mutation resulted in the creation of a new splice donor site which in turn resulted in a 24-bp (8 amino acids, RVRAGMCR) in-frame insertion. P-135 mutant mice thus carries a non-functional Itk protein with 8 amino acid insertion and may be useful in defining structural-functional relationships of Itk.
Publication Year: 2009
Publication Date: 2009-04-01
Language: en
Type: article
Indexed In: ['crossref']
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