Effect of 3,5-diethoxycarbonyl-1,4-dihydrocollidine on degradation of liver haem.
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Total RNA was isolated from mutants of Saccharomyces cerevisiae that lack active delta-aminolevulinate synthase and are therefore defective in heme biosynthesis. The RNAs were translated in the cell-free protein synthesis system from wheat germ, and the catalase T synthesized was isolated by immunoadsorption and polyacrylamide gel electrophoresis in the presence of dodecyl sulfate. Little or no catalase T product was detected with RNA from mutant cells grown in the absence of a heme precursor. As judged from in vitro translational capacity, RNA fractions from mutant cells grown in the presence of delta-aminolevulinate contained at least 10 times more catalase T mRNA than RNA from unsupplemented cells.
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The automatic determination of urinary ALA using a continuous flow methodology, is described. The amplification of the measured signal, with an adjusted range expander, allows to reduce the sample volume and to decrease drastically the effects of interfering substances. The results are compared with the reference method of Davis and Andelman [11].