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人肿瘤坏死因子相关凋亡诱导配体具有特异性诱导多种肿瘤细胞凋亡而对正常细胞无毒性的特点,被认为是肿瘤凋亡疗法较有希望的候选药物。植物生物反应器在药用蛋白质生产方面具备较大优势。构建了可溶性人肿瘤坏死因子相关凋亡诱导配体基因的植物表达载体,进行本氏烟(Nicotiana benthamiana)瞬时表达,并对目的蛋白的体外活性进行检测。结果显示,本氏烟叶片中目的蛋白平均表达量为68.60 pg/mg TSP;在浓度为200 pg/m L时,对NCI-H460细胞株的抑制率为28.75%。研究表明,经密码子优化的编码基因能够在受体植物中表达出有活性的目的蛋白,为开展目的基因的稳定转化工作奠定了基础。
Human Tumor Necrosis Factor Related Apoptosis-inducing Ligands, which have the characteristics of inducing apoptosis of many kinds of tumor cells without toxicity to normal cells, are considered as promising candidates for tumor apoptosis therapy. Plant bioreactors have great advantages in the production of medicinal proteins. The plant expression vector of soluble human tumor necrosis factor-related apoptosis-inducing ligand gene was constructed and transiently expressed in Nicotiana benthamiana, and the in vitro activity of the target protein was tested. The results showed that the average expression level of the target protein in N. benthamiana leaves was 68.60 pg / mg TSP. At the concentration of 200 pg / mL, the inhibitory rate of NCI-H460 cells was 28.75%. Studies have shown that codon-optimized coding genes can express active protein in recipient plants, which lays the foundation for the stable transformation of target gene.