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Optimization protein productivity of human interleukin-2 through codon usage,gene copy number and intracellular tRNA concentration in CHO cells
Authors:Kua-Chun Ou  Chih-Yang Wang  Kuan-Ting Liu  Yi-Ling Chen  Yi-Chen Chen  Ming-Derg Lai  Meng-Chi Yen
Institution:1. Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, No.1, University Rd., Tainan 701, Taiwan, ROC;2. Institute of Basic Medical Science, College of Medicine, National Cheng Kung University, No.1, University Rd., Tainan 701, Taiwan, ROC;3. Department of Emergency Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical University, No.100, Tzyou 1st Road, Kaohsiung 807, Taiwan, ROC;4. Department of Senior Citizen Service Management, Chia-Nan University of Pharmacy and Science, 60, Erh-Jen Rd., Sec.1, Jen-Te, Tainan 717, Taiwan, ROC;5. Infectious Diseases and Signaling Research Center, National Cheng Kung University, No.1, University Rd., Tainan 701, Taiwan, ROC
Abstract: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.
Keywords:tRNA abundance  Codon usage bias  Gene copy number  Interleukin-2  DNA vaccine
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