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Published on: August 5, 2011
Cloning and expression of the cDNA coding for the erythrocyte isoenzyme of human acylphosphatase
T Fiaschi1, G Raugei, R Marzocchini
1Department of Biochemical Sciences, University of Florence, Italy.
Abstract:
Three independent cDNAs coding for the erythrocyte isoform of human acylphosphatase were isolated and characterized. All the clones were incomplete at the 5' end, but Northern blot analysis using the cDNA as a probe showed the presence of an unusually long mRNA 5'-untranslated region. The transcript was present in a variety of human cell lines of different origins, although at different levels. Southern blot analysis on DNA from different individuals revealed a simple hybridization pattern. Large amounts of pure enzyme with kinetic characteristics very similar to those of the native protein were expressed in E. coli.
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