2020年5月1日
Reconstitution of yeast translation elongation and termination in vitro utilizing CrPV IRES-containing mRNA
The Journal of Biochemistry
- ,
- ,
- ,
- 巻
- 167
- 号
- 5
- 開始ページ
- 441
- 終了ページ
- 450
- 記述言語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1093/jb/mvaa021
- 出版者・発行元
- Oxford University Press (OUP)
<title>Abstract</title>
We developed an in vitro translation system from yeast, reconstituted with purified translation elongation and termination factors and programmed by CrPV IGR IRES-containing mRNA, which functions in the absence of initiation factors. The system is capable of synthesizing the active reporter protein, nanoLuciferase, with a molecular weight of 19 kDa. The protein synthesis by the system is appropriately regulated by controlling its composition, including translation factors, amino acids and antibiotics. We found that a high eEF1A concentration relative to the ribosome concentration is critically required for efficient IRES-mediated translation initiation, to ensure its dominance over IRES-independent random internal translation initiation.
We developed an in vitro translation system from yeast, reconstituted with purified translation elongation and termination factors and programmed by CrPV IGR IRES-containing mRNA, which functions in the absence of initiation factors. The system is capable of synthesizing the active reporter protein, nanoLuciferase, with a molecular weight of 19 kDa. The protein synthesis by the system is appropriately regulated by controlling its composition, including translation factors, amino acids and antibiotics. We found that a high eEF1A concentration relative to the ribosome concentration is critically required for efficient IRES-mediated translation initiation, to ensure its dominance over IRES-independent random internal translation initiation.
- リンク情報
- ID情報
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- DOI : 10.1093/jb/mvaa021
- ISSN : 0021-924X
- eISSN : 1756-2651