Figure 2.
The insertion changes expression of a reporter gene and creates predicted transcription factor binding sites. A. Chromatogram of the ZRS603ins13 (NG_009240.1:g.106934_106935ins13) insertion mutation. Sequencing a cloned allele of the ZRS from patient III/1 shows an insertion of 13 bp compared to the wildtype sequence represented by the sequence from an unaffected sibling (III/2). B. ZRS E11.5 mouse transgenic assay of human wtZRS showing posterior LacZ expression . C. ZRS E11.5 mouse transgenic assay of ZRS603ins13 showing strong posterior and ectopic anterior LacZ expression. D. The 13bp insertion, in bold, creates binding sites for multiple limb-associated transcription factors. Grey bars indicate the location of the TFBS.