Abstract
The main problem in studying mammalian selenocysteine-containing proteins is that the proteins are difficult to obtain in a recombinant form because the amino acid selenocysteine (Sec), which is their component, is encoded by TGA, which is one of the stop codons. When only the open reading frame of a target protein is cloned in a plasmid, translation is prematurely terminated at the TGA codon. An intricate natural mechanism allows the codon to be recognized as a selenocysteine codon and involves various cis- and trans-acting factors, such as the selenocysteine insertion sequence (SECIS), mRNA secondary structure, selenocysteine tRNA Sec-tRNA[Ser]Sec, SECIS-binding protein 2 (SBP2), selenocysteine-specific elongation factor EFsec, and others. Generation of recombinant selenoproteins in preparative amounts directly depends on the expression levels of the cis- and trans-acting transcription and translation factors to further complicate the problem, and cysteine homologs of selenoproteins are consequently used in many studies. Several methods designed to express mammalian selenoproteins in vitro are considered in the review.
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Abbreviations
- EFsec:
-
elongation factor of selenocysteine
- Sec:
-
selenocysteine
- SECIS:
-
selenocysteine insertion sequence
- SectRNA[Ser]Sec :
-
selenocysteine tRNA
- SBP2:
-
SECIS-binding protein 2
- SelA:
-
prokaryotic selenocysteine synthase
- SelB:
-
selenocysteine-specific elongation factor
- SelC:
-
selenocysteine-specific tRNA for thioredoxin reductase (TrxR)
- ORF:
-
open reading frame
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Original Russian Text © E.G. Varlamova, S.V. Novoselov, 2016, published in Molekulyarnaya Biologiya, 2016, Vol. 50, No. 1, pp. 44–50.
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Varlamova, E.G., Novoselov, S.V. Methods to biosynthesize mammalian selenocysteine-containing proteins in vitro. Mol Biol 50, 37–42 (2016). https://doi.org/10.1134/S0026893316010210
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DOI: https://doi.org/10.1134/S0026893316010210