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Selenocysteine tRNA[Ser]Sec, the central component of selenoprotein biosynthesis: Isolation, identification, modification, and sequencing

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Authors

Carlson, Bradley A.; Lee, Byeong Jae; Tsuji, Petra A.; Copeland, Paul R.; Schweizer, Ulrich; Gladyshev, Vadim N.; Hatfield, Dolph L.

Issue Date
2018-01
Publisher
Humana Press, Inc.
Citation
Methods in molecular biology (Clifton, N.J.), Vol.1661, pp.43-60
Abstract
© 2018, Springer Science+Business Media LLC.The selenocysteine (Sec) tRNA[Ser]Sec population consists of two isoforms that differ from each other by a single 2′-O-methylribosyl moiety at position 34 (Um34). These two isoforms, which are encoded in a single gene, Trsp, and modified posttranscriptionally, are involved individually in the synthesis of two subclasses of selenoproteins, designated housekeeping and stress-related selenoproteins. Techniques used in obtaining these isoforms for their characterization include extraction of RNA from mammalian cells and tissues, purifying the tRNA[Ser]Sec population by one or more procedures, and finally resolving the two isoforms from each other. Since some of the older techniques for isolating tRNA[Ser]Sec and resolving the isoforms are used in only a few laboratories, these procedures will be discussed briefly and references provided for more detailed information, while the more recently developed procedures are discussed in detail. In addition, a novel technique that was developed in sequencing tRNA[Ser]Sec for identifying their occurrence in other organisms is also presented.
ISSN
1064-3745
URI
https://hdl.handle.net/10371/203819
DOI
https://doi.org/10.1007/978-1-4939-7258-6_4
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