Production of recombinant proteins in plant cells

Gerasimova, S. V.; Smirnova, Olga; Кочетов, А. В.; Shumny, V. K.

doi:10.1134/s1021443716010076

Cited by 9 publications

(4 citation statements)

References 86 publications

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“…In addition, adventitious agents are absent in plants. Therefore, using a plant platform can decrease the risk of pathogen contamination [13]. Several plants, such as alfalfa, soybean, lettuce, potato, spinach, Arabidopsis, and tobacco, have been used to produce recombinant proteins [12][13][14][15].…”

Section: Introductionmentioning

confidence: 99%

“…Therefore, using a plant platform can decrease the risk of pathogen contamination [13]. Several plants, such as alfalfa, soybean, lettuce, potato, spinach, Arabidopsis, and tobacco, have been used to produce recombinant proteins [12][13][14][15]. Among these plants, tobacco has several benefits, such as well-established transformation protocols, high biomass, scale-up capacity, its role as a non-food crop, prolific seed production, and year-round growth.…”

Section: Introductionmentioning

confidence: 99%

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Recombinant Human Dentin Matrix Protein 1 (hDMP1) Expressed in Nicotiana benthamiana Potentially Induces Osteogenic Differentiation

Ahmad

Kaewpungsup

Khorattanakulchai

et al. 2019

Plants

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Inductive molecules are critical components for successful bone tissue engineering. Dentin matrix protein-1 (DMP1), a non-collagenous protein in the bone matrix, has been shown to play roles in osteogenic differentiation and phosphate homeostasis. This study aimed to produce recombinant human dentin matrix protein-1 (hDMP1) in Nicotiana benthamiana and investigated the ability of this plant-produced DMP1 to induce osteogenesis in human periodontal ligament stem cells (hPDLSCs). The hDMP1 gene was cloned into the geminiviral vector for transient expression in N. benthamiana. We found that hDMP1 was transiently expressed in N. benthamiana leaves and could be purified by ammonium sulphate precipitation followed by nickel affinity chromatography. The effects of hDMP1 on the induction of cell proliferation and osteogenic differentiation were investigated. The results indicated that plant-produced hDMP1 could induce the cell proliferation of hPDLSCs and increase the expression levels of osteogenic genes, including osterix (OSX), type I collagen (COL1), bone morphogenetic protein-2 (BMP2), and Wnt3a. Moreover, the plant-produced hDMP1 promoted calcium deposition in hPDLSCs as determined by alizarin red S staining. In conclusion, our results indicated that plant-produced hDMP1 could induce osteogenic differentiation in hPDLSCs and could potentially be used as a bone inducer in bone tissue engineering.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Recombinant Human Dentin Matrix Protein 1 (hDMP1) Expressed in Nicotiana benthamiana Potentially Induces Osteogenic Differentiation

Ahmad

Kaewpungsup

Khorattanakulchai

et al. 2019

Plants

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“…The plant system being presently developed for recombinant protein expression that is diverse in different technologies of gene transfer, expression strategies and methods of subsequent target protein extraction. The stable transformation (transgenic plant) and transient expression have been developed to express the protein in plant system (Demain et al, 2009;Gerasimova et al, 2016;Zagorskaya et al 2017).…”

Section: Plant Systemmentioning

confidence: 99%

“…Among the advantages, the most important is the time consumed for the entire expression, only approximately 2-12 days. Several plants have been developed to express the recombinant protein, including alfalfa, soybean, lettuce, potato, spinach, arabidopsis, and tobacco (Cao et al, 2017;Gerasimova et al, 2016). Tobacco has significant benefit, including established protocols of transformation, high yield expression, accessibility to scale up the production, non-consumption food, growth at any-time (Karg et al, 2009;Twyman et al, 2003).…”

Section: Transient Expressionmentioning

confidence: 99%

Expression of human dentin matrix protein 1 in escherichia coli and nicotiana benthamiana and their effect on proliferation and osteogenic differentiation of human periodontal ligament stem cells

Ahmad

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This study aimed to express recombinant protein human Dentin Matrix Protein 1 (hDMP1) in Escherichia coli and Nicotiana benthamiana and investigated the osteogenic differentiation effects in human Periodontal Ligament Stem Cells (hPDLSCs). The gene encoding hDMP1 was ligated into the pET22b and pBY2R2e expression vectors. Recombinant hDMP1 protein has been productively expressed in E. coli and N. benthamiana. E. coli-produced hDMP1 resulted in 60 kDa and 100, and the optimal condition to produce highly expression was demonstrated at 0.5 mM IPTG and incubation at 37oC for 6 hr. N. benthamiana-produced hDMP1 protein showed in a 100 kDa that indicated the presence of post-translational modification, and the highest-level expression was obtained at 2 dpi and OD600 0.4. Recombinant hDMP1 could induce ALP, BMP2, CBFA1, OSX, OPN, and WNT3a osteogenic marker genes and calcium deposition in hPDLSCc. In conclusion, the results indicated that recombinant hDMP1 could induce osteogenic differentiation in hPDLSCs. However, N. benthamiana-produced hDMP1 could induce osteogenic differentiation and calcium deposition better than E. coli-produced hDMP1. Therefore, N. benthamiana-produced hDMP1 has potential to use as an inductive molecule in bone tissue�engineering.

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Plant Molecular Farming for the Bulk Production of Industrial Enzymes

Basaran

2024

Concepts and Strategies in Plant Sciences

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Production of recombinant proteins in plant cells

Cited by 9 publications

References 86 publications

Recombinant Human Dentin Matrix Protein 1 (hDMP1) Expressed in Nicotiana benthamiana Potentially Induces Osteogenic Differentiation

Recombinant Human Dentin Matrix Protein 1 (hDMP1) Expressed in Nicotiana benthamiana Potentially Induces Osteogenic Differentiation

Expression of human dentin matrix protein 1 in escherichia coli and nicotiana benthamiana and their effect on proliferation and osteogenic differentiation of human periodontal ligament stem cells

Plant Molecular Farming for the Bulk Production of Industrial Enzymes

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