Development ofTomato bushy stunt virus-based vectors for fusion and non-fusion expression of heterologous proteins in an alternative hostNicotiana excelsiana
Development ofTomato bushy stunt virus-based vectors for fusion and non-fusion expression of heterologous proteins in an alternative hostNicotiana excelsiana
复制标题
开发基于番茄浓密矮化病毒的载体，用于在替代宿主烟草中融合和非融合表达异源蛋白。
DOI：
10.1007/s00253-020-10837-z
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发表时间：
2020-08-24
影响因子：
5
通讯作者：
Wang，Sheng
中科院分区：
文献类型：
--
作者：
Zhang，Xiqian;Ding，Xiangzhen;Wang，Sheng
摘要基于植物病毒的表达系统是生产临床和工业上有用的重组蛋白的另一种表达平台。然而，由于缺乏具有商业潜力的病毒载体，迫切需要设计和开发新的、通用的、高效的植物病毒载体。番茄丛矮病毒（tomato bushstunt virus， TBSV）的基因组为植物中产生异源蛋白的载体提供了一种有吸引力的替代方法。在此，我们开发了一套新的融合和非融合TBSV-CP替代载体，为表达植物中感兴趣的蛋白质提供了更灵活和高效的工具。本研究选用一种替代烟草植物excelsiana作为新构建的TBSV载体的宿主，因为报道了常用的benthamiannicotiana宿主与TBSV编码P19蛋白的表达相关的不良坏死效应。结果表明，TBSV载体引起无症状感染，报告基因inN过表达。excelsianaleaves，证明n。excelsian是tbsv介导的异源基因表达的理想寄主植物。此外，一种TBSV非融合载体daaug在并排比较中显示出与基于TMV和基于pvx的载体相似的报告蛋白积累水平，并且比先前开发的TBSV载体提供了更灵活的方面。总之，我们新开发的TBSV表达系统为植物病毒表达载体家族增加了一个新成员，同时为植物中产生感兴趣的蛋白质提供了一种灵活而高效的方法。•基于tbv的瞬时表达系统得到了显著改进。•利用棘球孢作为寄主植物，避免了病毒P19蛋白引起的坏死效应。•非融合载体的表达水平与迄今为止报道的最有效的病毒载体相似。
AbstractPlant virus-based expression systems are an alternative expression platform for the production of clinically and industrially useful recombinant proteins. Nonetheless, due to a lack of viral vector with the commercial potentials, it is urgent to design and develop new, versatile, and efficient plant virus vectors. The genome ofTomato bushy stunt virus(TBSV) offers an attractive alternative to being modified as a vector for producing heterologous proteins in plants. Here, we developed a set of novel fusion and non-fusion TBSV-CP replacement vectors, which provide more flexible and efficient tools for expressing proteins of interest in plants. An alternative tobacco plant,Nicotiana excelsiana, was used in this study as a host for newly constructed TBSV vectors because the unwanted necrotic effects were reported on the commonly usedNicotiana benthamianahost associated with expression of TBSV-encoded P19 protein. The data showed that TBSV vectors caused a symptomless infection and overexpressed reporter gene inN. excelsianaleaves, demonstrating thatN. excelsianais an ideal host plant for TBSV-mediated heterologous gene expression.Moreover, a TBSV non-fusion vector, dAUG, shows the similar accumulation level of reporter proteins to that of TMV- and PVX-based vectors in side-by-side comparison and provides more flexible aspects than the previously developed TBSV vectors. Collectively, our newly developed TBSV expression system adds a new member to the family of plant viral expression vectors and meanwhile offers a flexible and highly effective approach for producing proteins of interest in plants.Key points•The TBSV-based transient expression system has been significantly improved.•The necrotic effects caused by viral P19 protein were avoided by the usage of N. excelsiana as a host plant.•The expression level of the non-fusion vector was similar to the most effective virus vectors reported so far.