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. 2024 Nov 1;12(11):2505. doi: 10.3390/biomedicines12112505

Figure 1.

Figure 1

Generation of transgenic nZRS reporter newt. (a) PCR amplification of the newt (C. pyrrhogaster) ZRS enhancer from genomic DNA. F: forward degenerate primer; R: reverse degenerate primer. To rule out nonspecific PCR amplicons using degenerate primers, PCR reactions using a single primer (lane F or lane R) were used as controls. (b) Nucleotide sequence of the newt ZRS sequence, 819 bp (accession no. PP691624). Characterization of mouse binding sites identified in the newt nZRS in grey, vertebrate uncharacterized (UC) conserved sites in yellow, and urodele uncharacterized sites in green. Vertebrate ZRS alignments are available in Tables S1–S8 and Figure S3. Forward and reverse degenerate primer sets are shown on both ends of the sequence. Degenerate nucleotides indicated with bold. (c) Illustration of the transgene nZRS reporter plasmid. hmp: human minimal promoter; HS4: Chicken single core insulator. (d) Schematic summary of nZRS reporter transgenesis and screening.