Tg(Tagln-PLA2G2A*G29S)AMrj

The 7.2 kb transgene construct contains the promoter region of the mouse transgelin gene (Tgln; also called Sm22a) extending from -2126 to +4136 bp, including 2126 bp of 5' flanking sequence, the first exon, the first intron and the first 11 bp of the second intron, joined to a cDNA encoding a G29S mutant human phospholipase A2, group IIA (platelets, synovial fluid) (PLA2G2A) with a C-terminal V5 epitope tag and polyhistidine (6xHis) sequence. The glycine at amino acid position 29, in the calcium-binding loop of the protein, is essential for Ca2+ binding; its replacement by serine results in a catalytically inactive enzyme. Mutant line A is estimated by Southern blot analysis to carry more than 50 copies of the transgene. Analysis of liver, lung, skeletal muscle, heart and aorta of transgenic mice for transgene-derived PLA2G2A mRNA by reverse transcription- (RT-) PCR analysis and for the protein by western blot analysis demonstrated expression specifically in tissues enriched in smooth muscle, i.e., aorta, heart and lung. (J:34770, J:82809, J:209787)