Development of a Novel Cysteine Sulfinic Acid Decarboxylase Knockout Mouse: Dietary Taurine Reduces Neonatal Mortality
Figure 1
Analysis of the CSAD gene knockout. (a) Schematic diagram of pGT01xf insertion into the CSAD gene. Rectangles represent exons and open rectangles represent, untranslated regions. “A” shows the location of the VIS amplicon which is produced from the wild type chromosome and “B” shows the location of the β-geo amplicon which is produced from the gene trap vector inserted between exons 8 and 9. SA: splicing acceptor; pA: polyadenylation sequence; EV: EcoR V sites. (b) Southern analysis of genomic DNA digested with EcoR V and hybridized using [32P]-dCTP labeled CSAD cDNA. The wild type CSAD EcoR V fragment is 5.9 kb and that of disrupted fragment is 7.1 kb. “+/+”: WT; “+/−”: CSAD+/−; “−/−”: CSAD−/−. (c) PCR products (1038 bp and 682 bp) with VIS and β-geo primer sets. β-geo product is absent in WT and VIS PCR product is absent in CSAD−/−. Both PCR products are present in CSAD+/−. (d) Northern blot analysis of total RNA from liver with [32P]-dCTP labeled cDNA and -actin as a control probe. (e) Western blot analysis of kidney homogenates probed with an anti-CSAD antibody and detected with an alkaline phosphatase labeled goat anti-rabbit antibody. CSAD (51 kD) was not detected in CSAD−/− homogenates.
