人肿瘤坏死因子相关凋亡诱导配体基因植物表达载体的构建及瞬时表达

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。

Construction of Plant Expression Vector of shTRAIL and its Transient Expression

Human tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) induced apoptosis exhibits selective cytotoxicity, i.e. selectively induces apoptosis in numerous tumor cell lines while sparing normal cells, thus it has been regard as a promising candidate medicine of apoptosis-inducing therapy in malignancies. Plant bioreactor has been widely concerned due to its advantages in production of pharmaceutical proteins. Expression vector of soluble human TRAIL (shTRAIL) gene was constructed, and the Nicotiana benthamiana transient expression system was utilized in this paper. The results showed that the mean expression level of shTRAIL was 68.60 pg/mg TSP in N. benthamiana leaves, and the inhibition rate of NCI-H460 was 28.75% when treated with 200 pg/mL shTRAIL. This results indicated that the codon optimized gene could express target protein with biological activity in transgenic plants, and provide a basis for next stage of stable transformation of the target gene.