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검색결과 1,109건 처리시간 0.035초

Efficient Interleukin-21 Production by Optimization of Codon and Signal Peptide in Chinese Hamster Ovarian Cells

  • Cho, Hee Jun;Oh, Byung Moo;Kim, Jong-Tae;Lim, Jeewon;Park, Sang Yoon;Hwang, Yo Sep;Baek, Kyoung Eun;Kim, Bo-Yeon;Choi, Inpyo;Lee, Hee Gu
    • Journal of Microbiology and Biotechnology
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    • 제29권2호
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    • pp.304-310
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    • 2019
  • Interleukin-21 is a common ${\gamma}$-chain cytokine that controls the immune responses of B cells, T cells, and natural killer cells. Targeting IL-21 to strengthen the immune system is promising for the development of vaccines as well as anti-infection and anti-tumor therapies. However, the practical application of IL-21 is limited by the high production cost. In this study, we improved IL-21 production by codon optimization and selection of appropriate signal peptide in CHO-K1 cells. Codon-optimized or non-optimized human IL-21 was stably transfected into CHO-K1 cells. IL-21 expression was 10-fold higher for codon-optimized than non-optimized IL-21. We fused five different signal peptides to codon-optimized mature IL-21 and evaluated their effect on IL-21 production. The best result (a 3-fold increase) was obtained using a signal peptide derived from human azurocidin. Furthermore, codon-optimized IL-21 containing the azurocidin signal peptide promoted $IFN-{\gamma}$ secretion and STAT3 phosphorylation in NK-92 cells similar to codon-optimized IL-21 containing original signal peptide. Collectively, these results indicate that codon optimization and azurocidin signal peptides provide an efficient approach for the high-level production of IL-21 as a biopharmaceutical.

Pistacia weinmannifolia root exerts a protective role in ovalbumin-induced lung inflammation in a mouse allergic asthma model

  • Jae-Won Lee;Jae-Hong Min;Min-Gu Kim;Seong-Man Kim;Ok-Kyoung Kwon;Tae Kyu Oh;Jae Kyoung Lee;Tae Young Kim;Sang Woo Lee;Sangho Choi;Wan-Yi Li;Hyung Won Ryu;Kyung-Seop Ahn;Sei-Ryang Oh
    • International Journal of Molecular Medicine
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    • 제44권6호
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    • pp.2171-2180
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    • 2019
  • Pistacia weinmannifolia (Anacardiaceae) has been used in herbal medicine for the treatment of influenza, dysentery and enteritis in China. It was recently observed that P. weinmannifolia root extract (PWRE) exerts anti-inflammatory effects both in in vitro and in vivo models. Based on the results from previous studies, the present study investigated the protective effect of PWRE on airway inflammation and mucus hypersecretion. Treatment with PWRE significantly decreased the number of eosinophils and the levels of Th2 cytokines, such as interleukin (IL)-4, IL-5 and IL-13, in the bronchialveolar lavage fluid (BALF) of OVA-exposed mice. PWRE decreased the high serum levels of total and OVA-specific immunoglobulin E. PWRE also effectively inhibited the influx of inflammatory cells into the lung, as well as airway mucus hypersecretion. In addition, the increased level of monocyte chemoattractant protein-1 was significantly decreased with the PWRE treatment in the BALF of OVA-exposed mice and in lipopolysaccharide-stimulated RAW264.7 macrophages. These protective effects of PWRE on OVA-induced pulmonary inflammation were accompanied by the downregulation of mitogen associated protein kinases and nuclear factor-κB activation. Thus, the results from the present study indicate that PWRE could be valuable adjuvant for the treatment of asthma.

Pistacia weinmannifolia ameliorates cigarette smoke and lipopolysaccharide-induced pulmonary inflammation by inhibiting interleukin-8 production and NF-κB activation

  • Jae-Won Lee;Hyung Won Ryu;Su Ui Lee;Min-Gu Kim;Ok-Kyoung Kwon;Mun Ok Kim;Tae Kyu Oh;Jae Kyoung Lee;Tae Young Kim;Sang Woo Lee;Sangho Choi;Wan-Yi Li;Kyung-Seop Ahn;Sei-Ryang Oh
    • International Journal of Molecular Medicine
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    • 제44권3호
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    • pp.949-959
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    • 2019
  • Pistacia weinmannifolia (PW) has been used in traditional Chinese medicine to treat headaches, dysentery, enteritis and influenza. However, PW has not been known for treating respiratory inflammatory diseases, including chronic obstructive pulmonary disease (COPD). The present in vitro analysis confirmed that PW root extract (PWRE) exerts anti-inflammatory effects in phorbol myristate acetate- or tumor necrosis factor α (TNF-α)-stimulated human lung epithelial NCI-H292 cells by attenuating the expression of interleukin (IL)-8, IL-6 and Mucin A5 (MUC5AC), which are closely associated with the pulmonary inflammatory response in the pathogenesis of COPD. Thus, the aim of the present study was to evaluate the protective effect of PWRE on pulmonary inflammation induced by cigarette smoke (CS) and lipopolysaccharide (LPS). Treatment with PWRE significantly reduced the quantity of neutrophils and the levels of inflammatory molecules and toxic molecules, including tumor TNF-α, IL-6, IL-8, monocyte chemoattractant protein-1, neutrophil elastase and reactive oxygen species, in the bronchoalveolar lavage fluid of mice with CS- and LPS-induced pulmonary inflammation. PWRE also attenuated the influx of inflammatory cells in the lung tissues. Furthermore, PWRE downregulated the activation of nuclear factor-κB and the expression of phosphodiesterase 4 in the lung tissues. Therefore, these findings suggest that PWRE may be a valuable adjuvant treatment for COPD.

Genome-wide Drug-induced Haploinsufficiency Screening of Fission Yeast for Identification of Hydrazinocurcumin Targets

  • Baek, Seung-Tae;Kim, Dong-Uk;Han, Sang-Jo;Woo, Im-Sun;Nam, Mi-Young;Kim, Li-La;Heo, Kyung-Sun;Lee, Hye-Mi;Hwang, Hye-Rim;Choi, Shin-Jung;Won, Mi-Sun;Lee, Min-Ho;Park, Song-Kyu;Lee, Sung-Hou;Kwon, Ho-Jeong;Maeng, Pil-Jae;Park, Hee-Moon;Park, Young-Woo;Kim, Dong-Sup;Hoe, Kwang-Lae
    • Journal of Microbiology and Biotechnology
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    • 제18권2호
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    • pp.263-269
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    • 2008
  • Hydrazinocurcumin (HC), a synthetic derivative of curcumin, has been reported to inhibit angiogenesis via unknown mechanisms. Understanding the molecular mechanisms of the drug's action is important for the development of improved compounds with better pharmacological properties. A genome-wide drug-induced haploinsufficiency screening of fission yeast gene deletion mutants has been applied to identify drug targets of HC. As a first step, the 50% inhibition concentration $(IC_{50})$ of HC was determined to be $2.2{\mu}M$. The initial screening of 4,158 mutants in 384-well plates using robotics was performed at concentrations of 2, 3, and $4{\mu}M$. A second screening was performed to detect sensitivity to HC on the plates. The first screening revealed 178 candidates, and the second screening resulted in 13 candidates, following the elimination of 165 false positives. Final filtering of the condition-dependent haploinsufficient genes gave eight target genes. Analysis of the specific targets of HC has shown that they are related to septum formation and the general transcription processes, which may be related to histone acetyltransferase. The target mutants showed 65% growth inhibition in response to HC compared with wild-type controls, as shown by liquid culture assay.

Transgenic Sweetpotato Plants Expressing NDP Kinase 2

  • Lim, Soon;Yang, Kyoung-Sil;Park, Eun-Joon;Kwon, Suk-Yoon;Yun, Dae-Jin;Paek, Kee-Yoeup;Kwak, Sang-Soo;Lee, Haeng-Soon
    • 한국식물생명공학회:학술대회논문집
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    • 한국식물생명공학회 2005년도 춘계학술대회 및 국제심포지움 초록집
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    • pp.78-78
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    • 2005
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Expression of diligent protein and Pinoresinol/Lariciresinol reductase genes of forsythia in transgenic potatoes

  • Chuong, Tran-Van;Kim, Hyun-Soon;Park, Ji-Young;Joung, Jae-Youl;Youm, Jung-Won;Jeon, Jae-Heung
    • Plant Resources
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    • 제4권3호
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    • pp.181-188
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    • 2001
  • We tried to introduce two forsythia genes related in lignan biosynthesis, dirigent protein and pinoresinol/lariciresinol (Ph) reductase, into potatoes for accumulation of lignans in transgenic potatoes. We made binary vectors overexpressing dirigent protein gene and P/L reductase gene driven by a CaMV35S promoter and transformed into potatoes via Agrobacterium mediated transformation. And in order to control the metabolic flux of lignan biosynthesis pathway, we tried to inhibit chalcone synthase genes of potatoes by antisense inhibition technique also. We tried to use PCR screening method for selection of transgenic plants of different vectors. We tried to determine and compare lignan contents from different transgenic potato lines.

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