ArticleBMC biotechnology2009
High-level HIV-1 Nef transient expression in Nicotiana benthamiana using the P19 gene silencing suppressor protein of Artichoke Mottled Crinckle Virus.
Article in BMC biotechnology, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers.
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40 citing papers in PubMed, 80 citations in OpenAlex.
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- Nicotiana benthamiana's Responses to Agroinfiltration, a Treasure Grove of New Avenues to Improve Protein Yields in Plant Molecular Farming.Plant biotechnology journal · 2026Review
- Developing downstream processes for the purification of recombinant proteins and small molecules from Nicotiana benthamiana biomass.Plant biotechnology journal · 2026Review
- Transient expression of human interleukin-15 inPlant biotechnology (Tokyo, Japan) · 2025Article
- Recovery of RNA-dependent RNA polymerase 6 gene-knockout phenotypes in Nicotiana benthamiana via in vivo generation of inverted repeat construct of the trans-acting short interference RNA3 sequence.The Plant journal : for cell and molecular biology · 2025Article
- Rapid and Cost-Effective Diagnostic Blot Assays Based on the Use of Plant-Produced Recombinant Antigens: Lessons Learned from the SARS-CoV-2 RBD Antigen.International journal of molecular sciences · 2025Article
- Plant-related quality attributes affecting FcγRIIIa binding: affinity chromatography analysis of rituximab glycovariants fromFrontiers in plant science · 2025Article
- Plant production of recombinant antigens containing the receptor binding domain (RBD) of two SARS-CoV-2 variants.Biotechnology letters · 2024Article
- A Nitrogen-specific Interactome Analysis Sheds Light on the Role of the SnRK1 and TOR Kinases in Plant Nitrogen Signaling.Molecular & cellular proteomics : MCP · 2024Article
- Improving transient protein expression in agroinfiltrated Nicotiana benthamiana.The New phytologist · 2024Review
- Plant-Produced Viral Nanoparticles as a Functionalized Catalytic Support for Metabolic Engineering.Plants (Basel, Switzerland) · 2024Article
- Phase separation-based visualization of protein-protein interactions and kinase activities in plants.The Plant cell · 2023Article
- High-level transient production of a protease-resistant mutant form of human basic fibroblast growth factor inPlant biotechnology (Tokyo, Japan) · 2022Article
- UBP12 and UBP13 deubiquitinases destabilize the CRY2 blue light receptor to regulate Arabidopsis growth.Current biology : CB · 2022Article
- CRISPR/Cas9-mediated knockout of the DCL2 and DCL4 genes in Nicotiana benthamiana and its productivity of recombinant proteins.Plant cell reports · 2022Article
- Production of two SARS-CoV-2 neutralizing antibodies with different potencies inFrontiers in plant science · 2022Article
- Harnessing a Transient Gene Expression System inPlants (Basel, Switzerland) · 2021Article
- Fluorescent Labeling and Confocal Microcopy of Plastids and Stromules.Methods in molecular biology (Clifton, N.J.) · 2021Article
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Authors and funding
10 authors at 3 institutions in 1 country.
Funding
Abstract
backgroundIn recent years, different HIV antigens have been successfully expressed in plants by either stable transformation or transient expression systems. Among HIV proteins, Nef is considered a promising target for the formulation of a multi-component vaccine due to its implication in the first steps of viral infection. Attempts to express Nef as a single protein product (not fused to a stabilizing protein) in transgenic plants resulted in disappointingly low yields (about 0.5% of total soluble protein). In this work we describe a transient expression system based on co-agroinfiltration of plant virus gene silencing suppressor proteins in Nicotiana benthamiana, followed by a two-step affinity purification protocol of plant-derived Nef.
resultsThe effect of three gene silencing viral suppressor proteins (P25 of Potato Virus X, P19 of either Artichoke Mottled Crinckle virus and Tomato Bushy Stunt virus) on Nef transient expression yield was evaluated. The P19 protein of Artichoke Mottled Crinckle virus (AMCV-P19) gave the highest expression yield in vacuum co-agroinfiltration experiments reaching 1.3% of total soluble protein, a level almost three times higher than that previously reported in stable transgenic plants. The high yield observed in the co-agroinfiltrated plants was correlated to a remarkable decrease of Nef-specific small interfering RNAs (siRNAs) indicating an effective modulation of RNA silencing mechanisms by AMCV-P19. Interestingly, we also showed that expression levels in top leaves of vacuum co-agroinfiltrated plants were noticeably reduced compared to bottom leaves. Moreover, purification of Nef from agroinfiltrated tissue was achieved by a two-step immobilized metal ion affinity chromatography protocol with yields of 250 ng/g of fresh tissue.
conclusionWe demonstrated that expression level of HIV-1 Nef in plant can be improved using a transient expression system enhanced by the AMCV-P19 gene silencing suppressor protein. Moreover, plant-derived Nef was purified, with enhanced yield, exploiting a two-step purification protocol. These results represent a first step towards the development of a plant-derived HIV vaccine.
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