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Fig. 6
Generation of adam12-KO zebrafish. (A) adam12 gene sequence targeted for deletion and genotype of adam12-KO zebrafish. An alternative stop codon was inserted immediately after the metalloprotease active site as a result of a two-nucleotide (TG) deletion. (B) Whole-body images and genotypic ratio at 24-hpf of F1 embryos obtained by intercrossing adam12 (+/-) fish. Scale bar: 200 µm. (C) Comparison of adam12 mRNA expression levels by semi-quantitative RT-PCR between adam12 (+/+) and (-/-) zebrafish. Primers were designed to amplify the full-length coding sequence of adam12. Total RNA samples were isolated from 24-hpf embryos. Reverse-transcribed cDNA samples at the original concentration and 1/3 and 1/9 dilutions were used as PCR template. Expression of the β-actin gene was used as a positive control. (D) Comparison of adam8a, 8b, 10a, 10b, 17a, 17b, 19a, and 19b expression levels between adam12 (+/+) and (-/-). Primers were designed to amplify partial sequences of each adam gene.