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重组人八聚体结合转录因子 4 在水稻悬浮细胞中的表达。

Expression of Recombinant Human Octamer-Binding Transcription Factor 4 in Rice Suspension Cells.

机构信息

Graduate School of Biotechnology and Bioengineering, Yuan Ze University, 135 Yuan-Tung Road, Zhongli, Taoyuan County 320, Taiwan.

Taiwan Advance Bio-Pharmaceutical Inc., 12F, No. 25, Ln. 169, Kangning St, Xizhi Dist., New Taipei City 221, Taiwan.

出版信息

Int J Mol Sci. 2021 Jan 30;22(3):1409. doi: 10.3390/ijms22031409.

Abstract

The rice cell suspension culture system is a good way to produce recombinant human proteins, owing to its high biosafety and low production cost. Human Octamer-binding Transcription Factor 4 (Oct4) is a fundamental transcription factor responsible for maintaining human pluripotent embryonic stem cells. Recombinant Oct4 protein has been used to induce pluripotent stem cells. In this study, recombinant Oct4 proteins are produced via a sugar starvation-inducible / promoter-signal peptide-based rice recombinant protein expression system. mRNAs accumulate in the transgenic rice suspension cells under sugar starvation. The Oct4 recombinant protein is detected in the transgenic rice suspension cells, and its highest yield is approximately 0.41% of total cellular soluble proteins after one day of sugar starvation. The rice cell-synthesized recombinant human Oct4 protein show DNA-binding activity in vitro, which implies that the protein structure is correct for enabling specific binding to the target DNA motif.

摘要

水稻细胞悬浮培养系统是生产重组人蛋白的一种很好的方法,因为它具有很高的生物安全性和较低的生产成本。人八聚体结合转录因子 4(Oct4)是一种基本的转录因子,负责维持人类多能胚胎干细胞。重组 Oct4 蛋白已被用于诱导多能干细胞。在这项研究中,通过糖饥饿诱导/启动子信号肽的水稻重组蛋白表达系统生产重组 Oct4 蛋白。在糖饥饿下,转基水稻悬浮细胞中 mRNAs 积累。在转基水稻悬浮细胞中检测到 Oct4 重组蛋白,在糖饥饿 1 天后,其最高产量约为总细胞可溶性蛋白的 0.41%。水稻细胞合成的重组人 Oct4 蛋白在体外显示出 DNA 结合活性,这意味着该蛋白的结构正确,能够与目标 DNA 模体特异性结合。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4767/7866794/70184bd5856a/ijms-22-01409-g001.jpg

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