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5′‐poly(A) sequence as an effective leader for translation in eukaryotic cell‐free systems
Author(s) -
Gudkov Anatoly T.,
Ozerova Maria V.,
Shiryaev Vyacheslav M.,
Spirin Alexander S.
Publication year - 2005
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20525
Subject(s) - luciferase , translation (biology) , heterologous , cell free protein synthesis , green fluorescent protein , eukaryotic translation , sequence (biology) , protein biosynthesis , cell free system , messenger rna , translation system , wheat germ , microbiology and biotechnology , chemistry , biochemistry , biology , gene , transfection , enzyme , in vitro
Poly(A) sequence of 25 adenylic residues placed immediately before the start codons of the green fluorescent protein (GFP) and firefly luciferase (Luc) mRNAs is shown to provide a high rate of translation of the heterologous messages in eukaryotic cell‐free translation systems. Also the poly(A) leader is found to provide the abolition of the inhibition of translation at excess mRNA concentrations. The possibility of the practical use of the constructs with the poly(A) leader for preparative protein production is demonstrated in the wheat germ continuous‐exchange cell‐free (CECF) translation system. © 2005 Wiley Periodicals, Inc.