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. 2004 Oct 14;561(Pt 3):777–791. doi: 10.1113/jphysiol.2004.070631

Figure 1. TG constructs and mRNA expression.

Figure 1

A, schematic representation of the TG constructs. Mutant TM cDNAs (either Ser229Glu or His276Asn) were linked to the α–MHC promoter and used to generate multiple lines of TG mice (Subramaniam et al. 1991). The human growth hormone polyadenylation site (hgh poly A) was placed downstream of either TM cDNA. The black boxes denote the α–MHC exons that encode the 5′-untranslated region. The construct was released using BamHI enzyme. B and C, RNA expression in α-TM229 and α-TM276 TG mice, respectively; 20 μg of total cardiac RNA was hybridized to 3′-radioactively labelled DNA probes specific for either TM (see the small diagram below the RNA gel) or GAPDH. TM probe is 320 bp in length and yields fragments of 277 bp for mutant and 262 bp for endogenous TM RNA after S1 nuclease treatment. The electrophoresed samples shown here contain both mutant and endogenous bands for α-TM229 samples and an endogenous band only for NTG hearts. tRNA was used as a negative control.