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- W1545986340 abstract "This chapter presents the author's experience in cloning the β-hexosaminidase genes based on a polysome immunopurification approach. In this procedure, metabolically active cells are cracked open and polysomes—the cell's protein synthesis machinery—are isolated. The polysome contains an attached polypeptide chain in the process of translation that can be used as a tag for isolation of the β-hexosaminidase messenger RNA (mRNA) using specific antibodies. The isolated polysomes can be dissociated and the recovered mRNA reverse transcribed in cDNA for cloning. A drawback in the approach was that the antibodies were made against the human enzyme. The approach was to prepare two hybridization probes— one was a β-hexosaminidase- “enriched” probe made from the highly purified polysomal mRNA preparation, and the other was a β-hexosaminidase-“depleted” probe made from the polysomal mRNA preparation with the β-hexosaminidase mRNA removed. In addition to the identification of mutations, the cloned β-hexosaminidase sequences were fundamental for the elucidation of important structural and functional features of the enzyme. The cloning of the β-hexosaminidase genes has stimulated great progress in understanding the defects underlying Tay-Sachs disease (TSD) and the molecular biology of the β-hexosaminidase system." @default.
- W1545986340 created "2016-06-24" @default.
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- W1545986340 date "2001-01-01" @default.
- W1545986340 modified "2023-10-16" @default.
- W1545986340 title "11. Cloning the β-hexosaminidase genes" @default.
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- W1545986340 doi "https://doi.org/10.1016/s0065-2660(01)44075-2" @default.
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