Generation and heredity of the Brca1
L63X allele. (A) Structure of rat Brca1 and oligonucleotides used. Top, exon–intron structure. Middle, a magnified view. Bottom, nucleotide sequences. Arrowheads, primers; G1 and G2, guide RNAs; P1 and P2, probes. Uppercase, deoxyribonucleotides; lowercase, ribonucleotides; bold and underlined, location of the c.188T>A (p.L63X) mutation. (B) Sequence of cloned PCR products from the founder rat Brca1 exon 4. Arrows, induced mutation. (C–E) Heredity of the Brca1
L63X allele. (C) Standard curve of the allele‐specific quantitative PCR. ΔC
t, difference in the threshold cycles for the two molecular species. (D) Content of the Brca1
L63X allele in the genomic DNA of offspring (n = 40) of Brca1
L63X/+ heterozygotes. Mean and SD (duplicate measurements). (E) Contingency table of observed and expected numbers of offspring with each genotype. (F) Representative photographs of uteri of pregnant dams at designated days postcoitum (dpc). Arrowheads, maldeveloping conceptuses. (G) Weight of individual conceptuses