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C3P3-G1: first generation of a eukaryotic artificial cytoplasmic expression system.

Nucleic Acids Res.. 2019-03; 
JaïsPhilippe H,DecrolyEtienne,JacquetEric,Le BoulchMarine,JaïsAurélien,Jean-JeanOlivier,EatonHeather,PonienPrishila,VerdierFréderique,CanardBruno,GoncalvesSergio,ChironStéphane,Le GallMaude,MayeuxPatrick,Shmulevitz
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Plasmid DNA Preparation … to normalize for transfection efficacy, cells were transfected with the pORF-eSEAP plasmid (InvivoGen, San Diego, CA, USA), which encodes for … in Sf9 cells (Spodoptera frugiperda 9). The resulting sequence was synthesized into the F8 donor plasmid (GenScript, Piscataway, NJ … Get A Quote
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摘要

Most eukaryotic expression systems make use of host-cell nuclear transcriptional and post-transcriptional machineries. Here, we present the first generation of the chimeric cytoplasmic capping-prone phage polymerase (C3P3-G1) expression system developed by biological engineering, which generates capped and polyadenylated transcripts in host-cell cytoplasm by means of two components. First, an artificial single-unit chimeric enzyme made by fusing an mRNA capping enzyme and a DNA-dependent RNA polymerase. Second, specific DNA templates designed to operate with the C3P3-G1 enzyme, which encode for the transcripts and their artificial polyadenylation. This system, which can potentially be adapted to any... More

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