Molecular Plant Biophysics and Biochemistry, Department of Molecular Biology, University of Salzburg, Billrothstr. 11, 5020 Salzburg, Austria.
Plant Cell Rep. 2012 Mar;31(3):561-71. doi: 10.1007/s00299-011-1199-3. Epub 2011 Dec 13.
An economic and cheap production of large amounts of recombinant allergenic proteins might become a prerequisite for the common use of microarray-based diagnostic allergy assays which allow a component-specific diagnosis. A molecular pharming strategy was applied to express the major allergen of Artemisia vulgaris pollen, Art v 1, in tobacco plants and tobacco cell cultures. The original Art v 1 with its endogenous signal peptide which directs Art v 1 to the secretory pathway, was expressed in transiently transformed tobacco leaves but was lost in stable transformed tobacco plants during the alternation of generations. Using a light-regulated promoter and "hiding" the recombinant Art v 1 in the ER succeeded in expression of Art v 1 over three generations of tobacco plants and in cell cultures generated from stable transformed plants. However, the amounts of the recombinant allergen were sufficient for analysis but not high enough to allow an economic production. Although molecular pharming has been shown to work well for the production of non-plant therapeutic proteins, it might be less efficient for closely related plant proteins.
大量生产重组过敏原蛋白的经济且廉价的方法可能成为基于微阵列的诊断性过敏检测的常用手段,这种方法可以进行成分特异性诊断。应用分子农业策略在烟草植物和烟草细胞培养物中表达普通豚草花粉的主要过敏原 Art v 1。天然的 Art v 1 带有将其导向分泌途径的内源性信号肽,在瞬时转化的烟草叶片中表达,但在世代更替过程中在稳定转化的烟草植物中丢失。使用光调控启动子并将重组的 Art v 1“隐藏”在 ER 中,成功地在三代烟草植物和从稳定转化的植物中生成的细胞培养物中表达了 Art v 1。然而,重组过敏原的量足以进行分析,但不足以进行经济生产。尽管分子农业已被证明在生产非植物治疗性蛋白方面效果良好,但对于亲缘关系密切的植物蛋白,其效率可能较低。