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    Please use this identifier to cite or link to this item: http://ir.cnu.edu.tw/handle/310902800/28638

    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
    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
    Appears in Collections:[老人服務事業管理系] 期刊論文

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