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Plant virus expression systems for transient production of recombinant allergens in Nicotiana benthamiana.

机译:植物病毒表达系统,用于在本氏烟草中瞬时产生重组变应原。

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In recent years, several studies have demonstrated the use of autonomously replicating plant viruses as vehicles to express a variety of therapeutic molecules of pharmaceutical interest. Plant virus vectors for expression of heterologous proteins in plants represent an attractive biotechnological tool to complement the conventional production of recombinant proteins in bacterial, fungal, or mammalian cells. Virus vectors are advantageous when high levels of gene expression are desired within a short time, although the instability of the foreign genes in the viral genome may present problems. Similar levels of foreign protein production in transgenic plants often are unattainable, in some cases because of the toxicity of the foreign protein. Now virus-based vectors are for the first time investigated as a means of producing recombinant allergens in plants. Several plant virus vectors have been developed for the expression of foreign proteins. Here, we describe the utilization of tobacco mosaic virus- and potato virus X-based vectors for the transient expression of plant allergens in Nicotiana benthamiana plants. One approach involves the inoculation of tobacco plants with infectious RNA transcribed in vitro from a cDNA copy of the recombinant viral genome. Another approach utilizes the transfection of whole plants from wounds inoculated with Agrobacterium tumefaciens containing cDNA copies of recombinant plus-sense RNA viruses.
机译:近年来,几项研究证明了使用自主复制的植物病毒作为媒介物来表达多种具有药物意义的治疗分子。用于在植物中表达异源蛋白的植物病毒载体代表了一种有吸引力的生物技术工具,可补充细菌,真菌或哺乳动物细胞中重组蛋白的常规生产。当在短时间内需要高水平的基因表达时,病毒载体是有利的,尽管病毒基因组中外源基因的不稳定性可能会带来问题。在某些情况下,由于外源蛋白质的毒性,通常无法获得转基因植物中相似水平的外源蛋白质生产。现在,首次研究了基于病毒的载体,作为在植物中产生重组变应原的一种手段。已经开发了几种用于表达外源蛋白质的植物病毒载体。在这里,我们描述了基于烟草花叶病毒和马铃薯X病毒的载体在本生烟草中植物变应原的瞬时表达中的利用。一种方法涉及用从重组病毒基因组的cDNA拷贝体外转录的感染性RNA接种烟草植物。另一种方法利用接种了根癌农杆菌的伤口转染整个植物,所述根癌农杆菌含有重组正义RNA病毒的cDNA拷贝。

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