A technique to increase protein yield in a rabbit reticulocyte lysate translation system.
Rabbit reticulocyte lysate (RRL) is a mammalian cell-free system for protein production. However, one of the limitations of this system is its low protein yield. Inclusion of recombinant virus proteins and specific viral structures on target mRNA could enhance protein production in RRL. Here we show that simultaneous addition of influenza A virus NS1 protein and inclusion of the encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES) in the target mRNA facilitate translation initiation and increase protein yield over 10-fold, improving the translation capacity of RRL.
Institute for Molecular Medicine Finland (FIMM), University of Helsinki, Finland.
Maria
M
Anastasina
Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russian Federation.
Ilya
I
Terenin
Institute of Biotechnology, University of Helsinki, Finland.
Sarah J
SJ
Butcher
Institute for Molecular Medicine Finland (FIMM), University of Helsinki, Finland.
Denis E
DE
Kainov
D000818:Animals; D002474:Cell-Free System; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis; D012156:Reticulocytes / Q000378:metabolism*; D017361:Viral Nonstructural Proteins / Q000235:genetics
C085820:INS1 protein, influenza virus; D012333:RNA, Messenger; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins
EMCV IRES; cell-free translation; influenza A virus; non-structural protein; rabbit reticulocyte lysate