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Fig. 1. Targeted disruption of the Mili gene. (A) Schematic representation
of the wild-type allele, the targeting vector and the mutated alleles. The
numbered boxes (1-6) denote the 5'-end non-coding exon (exon 1) and the
five coding exons (exons 2-5). The targeting vector includes the
PGK-neo gene (neo) and the thymidine kinase gene (TK). (B)
Southern blot analysis of representative offspring from heterozygous mating.
The wild-type allele produces a 10.1 kb EcoRV product, while the
disrupted allele gives rise to a 5.5 kb band with the 5'-end
hybridization probe. (C) Western blot analysis of the MILI and MIWI proteins
from testes using antibody 26F, which recognizes both MILI and MIWI. Lysates
(10 µg of protein in each lane) of the testes were loaded on the gel. The
+/+, +/- and -/- designations indicate samples from wild-type, heterozygous
and homozygous mutant testes, respectively. The 293T cells that were
transfected with Mili- or Miwi-expressing plasmids and mock
plasmids are shown as the controls.
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