title: Identification of short untranslated regions that sufficiently enhance translation in high-quality wheat germ extract.
abstract: High-quality wheat germ extract (hqWGE) is very useful for the high-yield production of various types of protein. The most important key to high productivity is the design of mRNA templates. Although the design has been refined for straightforward and efficient translation in hqWGE, there is still room for improvement in untranslated regions (UTRs), especially the 3' UTR length, because a long, cumbersome 3' UTR is commonly used for translation enhancement. Here we examined some short viral 3' cap-independent translation enhancers (3' CITEs) to identify effective ones for efficient translation in hqWGE. We then combined the most effective 3' CITE and a 5' enhancer to further increase the translation efficiency. mRNA with the optimal short 3' and 5' UTRs, both of whose length was less than 150 nt, exhibited a productivity of 1.4 mg/mL in prolonged large-scale protein synthesis in hqWGE, which was comparable to that of control mRNA with a commonly-used long 3' UTR (∼1200 nt).
authors.affiliation: Proteo-Science Center, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime 790-8577, Japan. Electronic address: ogawa.atsushi.mf@ehime-u.ac.jp.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Ogawa
authors.affiliation: Proteo-Science Center, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime 790-8577, Japan.
authors.forename: Junichiro
authors.initials: J
authors.lastname: Tabuchi
authors.affiliation: Proteo-Science Center, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime 790-8577, Japan.
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Doi
mesh_terms: D010936:Plant Extracts / Q000737:chemistry / Q000378:metabolism*; D014176:Protein Biosynthesis; D011506:Proteins / Q000378:metabolism*; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism*; D014908:Triticum / Q000737:chemistry / Q000378:metabolism*
chemical_list: D010936:Plant Extracts; D011506:Proteins; D012333:RNA, Messenger
keywords: Cap-independent translation; Cell-free translation; Translation enhancer; Untranslated region; Wheat germ extract