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| 標題: | Optimization protein productivity of human interleukin-2 through codon usage, gene copy number and intracellular tRNA concentration in CHO cells |
| 作者: | Ou, Kua-Chun
Wang, Chih-Yang
Liu, Kuan-Ting
Chen, Yi-Ling
Chen, Yi-Chen
Lai, Ming-Derg
Yen, Meng-Chi |
| 貢獻者: | 老人服務事業管理系 |
| 關鍵字: | tRNA abundance
Codon usage bias
Gene copy number
Interleukin-2
DNA vaccine |
| 日期: | 2014-11 |
| 上傳時間: | 2015-05-06 21:23:26 (UTC+8) |
| 出版者: | Academic Press Inc Elsevier Science |
| 摘要: | Transfer RNA (tRNA) abundance is one of the critical factors for the enhancement of protein productivity in prokaryotic and eukaryotic hosts. Gene copy number of tRNA and tRNA codon usage bias are generally used to match tRNA abundance of protein-expressing hosts and to optimize the codons of recombinant proteins. Because sufficient concentration of intracellular tRNA and optimized codons of recombinant proteins enhanced translation efficiency, we hypothesized that sufficient supplement of host's tRNA improved protein productivity in mammalian cells. First, the small tRNA sequencing results of CHO-K1 cells showed moderate positive correlation with gene copy number and codon usage bias. Modification of human interleukin-2 (IL-2) through codons with high gene copy number and high codon usage bias (IL-2 HH, modified on Leu, Thr, Glu) significantly increased protein productivity in CHO-K1 cells. In contrast, modification through codons with relatively high gene copy number and low codon usage bias (IL-2 HL, modified on Ala, Thr, Val), or relatively low gene copy number and low codon usage bias (IL-2 LH, modified on Ala, Thr, Val) did not increase IL-2 productivity significantly. Furthermore, supplement of the alanine tRNA or threonine tRNA increased IL-2 productivity of IL-2 HL. In summary, we revealed a potential strategy to enhance productivity of recombinant proteins, which may be applied in production of protein drug or design of DNA vaccine. (C) 2014 Elsevier Inc. All rights reserved. |
| 關聯: | Biochemical and Biophysical Research Communications, v.454 n.2, pp.347-352 |
| 顯示於類別: | [高齡福祉養生管理系] 期刊論文
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