Figure 3
Production of the V1a2 knockout lineage. (a) Results of the heteroduplex mobility assay (HMA) analysis of all V1a2 KO F0 generation that survived 5Ā months of age. W indicates wild type. NC indicates negative control. (b) The gene sequence and amino acid sequence of the target site of the F0 generation, which had a particularly high mutagenesis rate and was selected for F1 generation production (one female and two males). The label number (#) of the individual matches the number of HMA analysis result in (a). Alignment results of the data obtained by sequence analysis of 10 clones of each fish are shown. The left and right TALEN recognition areas are highlighted in yellow. Ī indicates a base deletion, and plus indicates a base insertion. *indicates a stop codon. (c) As a result of sequence analysis of the target gene sites of all F1 fish that survived 5ā6Ā months post-hatch, three patterns of defective mutation and two patterns of base insertion were confirmed. (d) Breakdown of mutants of both alleles in all F1 fish sequenced. In 30 individuals, the genotype is still unknown, since clear sequence analysis results were not obtained.
