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• W296085402 abstract "This chapter discusses a cell-free system for polyadenylation using mouse B-cell extracts. The development of cell-free systems to study the initiation and regulation of transcription and the RNA processing reactions—splicing and polyadenylation—represents a major advance in the biochemical analysis of reactions. The chapter describes a detailed procedure for in vitro polyadenylation using cell-free protein extracts prepared from mouse cells of lymphoid origin. This system was primarily developed to study the posttranscriptional regulation of the mouse immunoglobulin M (IgM) locus. Polyadenylation of pre-RNA requires two independent steps. The first step is an endonucleolytic cleavage of the pre-RNA at a unique phosphate bond called the poly(A) site. After the cleavage approximately 200 adenosine residues are added to the 3´-OH of the upstream cleavage product. Sequences located both upstream and downstream of the poly(A) site are required for the reaction. The development of the in vitro polyadenylation system based on HeLa cell nuclear extract prompted several laboratories to study the reaction at the biochemical level." @default.
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• W296085402 date "1990-01-01" @default.
• W296085402 modified "2023-10-18" @default.
• W296085402 title "Cell-Free System for Polyadenylation Using Mouse B Cell Extracts" @default.
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• W296085402 doi "https://doi.org/10.1016/b978-0-12-442704-4.50012-8" @default.
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