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- W2386830534 abstract "MES23.5 cell line, which served as a useful tool for studying neurodegenerative disease, is a dopaminergic neuroblastoma derived from somatic cell fusion of rat embryonic mesencephalon cells and the murine neuroblastoma-glioma cells. But the origin of its tyrosine hydroxylase (TH) was unclear. A TH cDNA was cloned from MES23.5 cells by RT-PCR. The 1497 bp cDNA, encoding 498 amino acids, is completely identical to the mouse TH and shares 93% homology with rat TH. These results reveal that MES cells express mouse TH. The coding region of the cDNA was subcloned into a bacteria expression vector pGEX-4T-1, and the resulting TH-GST fusion protein was expressed at a high level in E. coli and purified via affinity chromatography. An improved method is used to analyze the activity of TH in vitro. The specific enzyme activity was 0.2698 μmol/min per mg of protein. The molecular mass of rTH—? ?6 000 was measured by SDS-polyacrylamide gel electrophoresis and the optimal pH of the enzymatic activity was 7.0. The activity was significantly inhibited by EDTA, and was markedly activated by Fe(Ⅱ)." @default.
- W2386830534 created "2016-06-24" @default.
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- W2386830534 date "2004-01-01" @default.
- W2386830534 modified "2023-09-25" @default.
- W2386830534 title "Identification and Activity Assay of the Tyrosine Hydroxylase in MES23.5 Cells" @default.
- W2386830534 hasPublicationYear "2004" @default.
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