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首页> 外文期刊>African Journal of Biotechnology >Expression of human soluble tumor necrosis factor (TNF)-related apoptosis-inducing ligand in transplastomic tobacco
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Expression of human soluble tumor necrosis factor (TNF)-related apoptosis-inducing ligand in transplastomic tobacco

机译:人可溶性肿瘤坏死因子相关的凋亡诱导配体在转基因烟草中的表达

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摘要

The soluble extracellular domain of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (sTRAIL) can, as the whole length?TRAIL?protein, bind with its receptors and specifically induce the apoptosis of cancer cells; therefore, it has been developed as a potential therapeutic agent for various cancer treatments. As it has become an attractive technology for foreign protein production, especially for production of biopharmaceuticals, chloroplast engineering was applied in this study to express human?sTRAIL?protein in tobacco. Two transplastomic lines were obtained. Southern blot showed that?sTRAIL?gene was inserted into the right site of the tobacco chloroplast genome. RT-PCR results also confirmed that the foreign gene is transcribed in both lines. However, western blot showed that only one line accumulated?sTRAIL?protein stably, while the other line lost the ability to accumulate this protein after several rounds of subcultures. The possible reason for this unexpected phenomenon is discussed.
机译:肿瘤坏死因子(TNF)相关的凋亡诱导配体(sTRAIL)的可溶性细胞外结构域可以与全长的TRAILβ蛋白结合,并与其受体结合,并特异性地诱导癌细胞的凋亡。因此,已经开发出其作为用于各种癌症治疗的潜在治疗剂。由于它已成为生产外国蛋白质,尤其是生产生物制药的一种有吸引力的技术,因此本研究应用叶绿体工程技术在烟草中表达人sTRAIL?蛋白。获得了两个转质体系。 Southern印迹表明,sTRAIL基因已插入烟草叶绿体基因组的正确位点。 RT-PCR结果也证实了外源基因在两个系中都被转录。然而,蛋白质印迹显示只有一系稳定地积累了sTRAILβ蛋白,而另一系在经过几轮亚培养后丧失了积累该蛋白的能力。讨论了此意外现象的可能原因。

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