5'-poly(A) sequence as an effective leader for translation in eukaryotic cell-free systems.
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.
Institute of Protein Research, Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russia.
Anatoly T
AT
Gudkov
Maria V
MV
Ozerova
Vyacheslav M
VM
Shiryaev
Alexander S
AS
Spirin
D020121:5' Untranslated Regions / Q000235:genetics / Q000502:physiology*; D000818:Animals; D001483:Base Sequence; D002474:Cell-Free System / Q000502:physiology*; D004742:Enhancer Elements, Genetic; D005057:Eukaryotic Cells / Q000378:metabolism*; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D013997:Time Factors
D020121:5' Untranslated Regions; D012333:RNA, Messenger