在 L.中高效表达和纯化人表皮生长因子
High Efficient Expression and Purification of Human Epidermal Growth Factor in L.
机构信息
State Key Laboratory of Crop Biology, Shandong Agricultural University, Tai'an 271018, China.
College of Pharmaceutical Sciences, Binzhou Medicial University, Yantai 264003, China.
出版信息
Int J Mol Sci. 2019 Apr 25;20(8):2045. doi: 10.3390/ijms20082045.
BACKGROUND
Human epidermal growth factor (hEGF) has drawn intense research attention due to its potential ability to promote healing of serious injuries, such as cuts, burns, and diabetic ulcers. Although hEGF displays prospective clinical value, the growth factor is restricted to the treatment of chronic diabetic ulcers because of its high production cost.
METHODS
Leguminous plant peanut ( L.) hairy roots contain relatively few toxic and harmful substances, and tested as an excellent production system for hEGF in our study. To explore the possibility of hEGF expression in peanut, overexpression hairy roots were obtained by infecting leaves with R1601.
RESULTS
The maximum transgenic hairy roots inducing rate was 82%. Protein purification and mass spectrometry assays showed that the protein expressed in peanut hairy roots was identified as hEGF. Furthermore, Methylthiazolyldiphenyl-tetrazolium bromide assay showed that hEGF promoted HL-7702 liver cells proliferation, which indicate that hEGF has biological activity and non-toxic on human cells.
CONCLUSION
Our results demonstrate the capacity of peanut hairy root cultures as a controlled, sustainable, and scalable production system that can be induced to produce valued human proteins, such as hEGF.
背景
由于人表皮生长因子（hEGF）具有促进严重创伤（如切割伤、烧伤和糖尿病溃疡）愈合的潜力，因此引起了人们的浓厚研究兴趣。尽管 hEGF 具有潜在的临床价值，但由于其生产成本高，该生长因子仅限于治疗慢性糖尿病溃疡。
方法
豆科植物花生（L.）毛状根含有相对较少的有毒有害物质，并且在我们的研究中被测试为 hEGF 的优秀生产系统。为了探索在花生中表达 hEGF 的可能性，我们用 R1601 感染叶片获得了过表达的毛状根。
结果
最大的转基因毛状根诱导率为 82%。蛋白纯化和质谱分析表明，在花生毛状根中表达的蛋白被鉴定为 hEGF。此外，噻唑蓝溴化二苯基四唑（MTT）比色法表明，hEGF 可促进 HL-7702 肝细胞增殖，表明 hEGF 对人细胞具有生物活性和无毒。
结论
我们的结果表明，花生毛状根培养物作为一种可控制、可持续和可扩展的生产系统，能够诱导产生有价值的人类蛋白，如 hEGF。
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