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Fig. 1. Targeted disruption of the mouse Dlgh1 gene. (A) The
design of the Dlgh1 targeting construct and the structure of the
mutant allele. The neo gene is inserted into a ClaI site, and is
flanked by 0.85 kb and 8.5 kb homologous sequences at the 5' and
3' sides (thick lines), respectively. The thin lines indicate sequences
derived from the pBluescript vector. The locations of the DNA probe used for
Southern blot analysis and the PCR primers used for genotyping are indicated.
Black boxes represent exons. Bam, BamHI; Cla, ClaI; Eco,
EcoRI; Xho, XhoI. (B) Southern blot analysis of tail
DNA from F3 progeny using EcoRI-digested genomic DNA. (C) PCR
genotyping of Dlgh1-/- mice. Competitive PCR was performed
on genomic tail DNA with three primers (see A): one common 3' primer
(R), a 5' Dlgh1-specific primer (F) and a 5' neo-specific
primer (N). DNA fragments of 503 bp and 273 bp are amplified from the
wild-type and mutant alleles, respectively. (D) Western blot analysis
of brain and kidney extracts from Dlgh1+/+,
Dlgh1+/- and Dlgh1-/- mice at E15.5.
Dlgh1 is not detected in the homozygous mutant mice, and the expression levels
of G3PDH are similar among the genotypes.
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