Development of an advanced polysome display system dependent on a specific protein-RNA motif interaction.
The ribosome display system is a very powerful too for in vitro screening of mRNAs that encode proteins (or peptides) with specific (known and unknown) functions. The ribosome display system depends on the stability of ribosome-mRNA complexes that have been achieved by the removal of a stop codon. An additional interaction we employed was between tandem fused MS2 coat protein (MSp) dimer and the RNA of its specific motif called a "C-variant" (or "Cv"). The MSp dimer and the Cv were placed at the N-terminal end of a nascent protein translated in vitro and the 5'-end of its mRNA, that might circularize the polysome complex and, as a result, further stabilize the ribosome-mRNA complex. The selectivity of mRNAs after selection was increased 2-fold by the introduction of the interaction between the MSp dimer and Cv.
Sawata
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SY
National Institute for Advanced Interdisciplinary Research, 1-1 Higashi, Tsukuba Science City, Ibaraki 305-8562, Japan.
Taira
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D001483:Base Sequence; D001665:Binding Sites; D036022:Capsid Proteins / Q000235:genetics* / Q000378:metabolism; D018388:Codon, Terminator; D005821:Genetic Techniques; D008957:Models, Genetic; D011132:Polyribosomes / Q000737:chemistry / Q000378:metabolism*; D014176:Protein Biosynthesis; D011506:Proteins / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis* / Q000378:metabolism; D012367:RNA, Viral / Q000737:chemistry* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism
D036022:Capsid Proteins; D018388:Codon, Terminator; D011506:Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; D016601:RNA-Binding Proteins