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Utilizing Plant Cell Suspension Platform BY-2 for the Production of Beta-1,3-Glucanase Enzyme
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Reza Mohammadzadeh    , Mostsfa Motallebi * , Zahra Moghaddassi jahromi |
| Plant Molecular Biotechnology Department, National Institute of Genetic Engineering and Biotechnology , motalebi@nigeb.ac.ir |
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Abstract: (147 Views) |
Due to the increasing demand for animal-free products, plant cells are becoming a suitable platform for producing recombinant proteins of interest that are not derived from animal clls. Plant cells have advantages such as low maintenance cost, ease of separation and purification of the produced products, and independent cultivation from weather conditions, soil quality, seasons, day length, and weather conditions. In this study, BY-2 plant cells were used as a platform for producing recombinant proteins. BY-2 cells have a high cell division rate, which is an important feature of these cells. To optimize the transfer and expression of genes in BY-2 cells, the gus reporter gene was transferred to these cells. Expression of this gene leads to a measurable phenotype. To confirm the presence of the gus gene in BY-2 cells, the expected 521bp fragment was observed using PCR. Additionally, to further confirm the biochemical expression of GUS in BY-2 cells, cell blue staining was observed. To express the beta-1,3-glucanase enzyme, the intron containing bgnI gene from the T. virens fungal strain was transferred to BY-2 cells using the pBI121GUS- dual vector. The expression of the protein in BY-2 cells was confirmed by SDS-PAGE analysis. The activity of the beta-1,3-glucanase enzyme (BgnI) was determined using the dinitrosalicylic acid (DNS) method to measure the amount of reducing sugars produced by the enzyme. The results showed that the transgenic BY-2 cells were capable of expressing active beta-1,3-glucanase enzyme.
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| Keywords: beta-1, 3-glucanase enzyme, recombinant proteins, bgnI gene, BY-2 cell, glucan. |
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Type of Study: Research |
Subject:
Plant
Received: 2023/06/19 | Accepted: 2023/09/8 | Published: 2024/03/14
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