Figure 1.

Design of hSOD1^mus gene constructs and generation of tg mice. (A) Schematic representation of constructs used in creating hSOD1^mus mice. The skeletal muscle-specific α-actin promoter was used to drive expression of human wild-type or mSOD1. The human wild-type cDNA is shown. Two different point mutations were introduced by site-specific mutagenesis to convert codon 37 from a glycine to an agrinine or codon 93 from a glycine to an alanine. The probe used for Southern analysis spanned the entire chicken α_sk actin promoter and coding region of hSOD1. (B) Southern blot analysis of hSOD1^mus transgene copy number in different tg mouse lines. NcoI digestion of mouse tail genomic DNA yields a 2.5 kb fragment in tg mice. Positive controls/copy number standards were digested tail DNA from non-tg mice spiked with various amounts of ApaLI-digested construct. The negative control (0) was non-spiked DNA from non-tg mice. (C) qPCR analysis of hSOD1 transgene copy number in representative hSOD1^mus tg G37R mice (lines 25, 59 and 73), G93A mice (lines 98 and 112) and wild-type mice (lines 131 and 224). Bars represent mean ± SEM and are the results of triplicate experiments.