Development of Tomato bushy stunt virus -based vectors for fusion and non-fusion expression of heterologous proteins in
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APPLIED GENETICS AND MOLECULAR BIOTECHNOLOGY
Development of Tomato bushy stunt virus-based vectors for fusion and non-fusion expression of heterologous proteins in an alternative host Nicotiana excelsiana Xiqian Zhang 1 & Xiangzhen Ding 1,2 & Zhiying Li 1,2 & Sheng Wang 1,2,3 Received: 18 June 2020 / Revised: 22 July 2020 / Accepted: 14 August 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Plant 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 of Tomato 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 nonfusion 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 used Nicotiana benthamiana host associated with expression of TBSV-encoded P19 protein. The data showed that TBSV vectors caused a symptomless infection and overexpressed reporter gene in N. excelsiana leaves, demonstrating that N. excelsiana is 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. Keywords Tomato bushy stunt virus . Viral vector . Nicotiana excelsiana . Non-fusion vector . Superexpression
Introduction Plant virus-based vectors have been regarded as a potential tool for the production of proteins of interest with a limited * Sheng Wang [email protected] 1
Key Laboratory of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, Yinchuan 750021, China
2
School of Life Science, Ningxia University, 539 W Helanshan Road, Yinchuan 750021, China
3
Key Laboratory of Modern Molecular Breeding for Dominant and Special Crops in Ningxia, Yinchuan 750021, China
cost (Streatfield 2007). An outstanding advantage of the viral vector sys
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