Figure 1

Construction of TALEN-based mutants of the DH-PBAN and DHR genes.

Schematic representations of the genes (top) and cDNA structures (bottom) of the DH-PBAN (A) and DHR (B) mutants. Shaded boxes and lines represent exons and introns, respectively. The cDNA of DH-PBAN encodes a signal sequence (S.S), DH, PBAN and α-, β- and γ-SGNPs, which carry the 5′ and 3′ untranslated regions (light-grey boxes on left and right sides, respectively). The cDNA of DHR consists of seven transmembrane domains (1–7), which carry the 5′ and 3′ untranslated regions (light-grey boxes on left and right sides, respectively). The FXPRLa and transmembrane domains are indicated using blue boxes. Orange triangles represent the TALEN binding sites. The sizes of exons and introns (in bp) are indicated by scales in each map. (C,D) The sequences of the TALEN target sites are indicated by orange boxes. Partial coding sequences corresponding to the DH N-terminus and transmembrane domain 1 of DHR are indicated using blue boxes. The deleted bases in spacer regions of each mutant are indicated by hyphens and identical bases are indicated by asterisks. The ΔDHP33, ΔDHP531 and ΔDHR96 are truncated proteins, which encode 9, 26 and 28 amino acids, respectively. Eight amino acids were missing from DHR111, which led to the production of a defective truncated protein in the extracellular domain at the N-terminus of DHR.