Comparison of Transient HDA19 Protein Expression in Leaves of Nicotiana tabacum and Nicotiana bentamiana

Document Type : Research Paper

Authors

1 Ph.D. Student, Department of Plant Breeding & Biotechnology, Sari Agricultural Sciences and Natural Resources University (SANRU), Sari, Iran

2 Associate Professor, Department of Plant Breeding & Biotechnology, Sari Agricultural Sciences and Natural Resources University (SANRU), Sari, Iran

3 Institute of Biochemical Plant Pathology, Helmholtz Zentrum München, German Research Center for Environmental Health, Neuherberg, 85764, Germany

4 Assistant Professor, Department of Plant Breeding & Biotechnology, Sari Agricultural Sciences and Natural Resources University (SANRU), Sari, Iran

Abstract

Recently, transient gene expression has been developed to provide a more rapid means of assessing plant tissues as a protein production platform without the labor-intensive and time-consuming process of generating stably transformed transgenic plants. This study reports the expression of HDA19 gene in two species of tobacco plants (Nicotiana tabacum and Nicotiana bentamiana) by means of transient transformation. Specific primers were designed and used for PCR amplification and cloning of HDA19 gene in the plant expression vector pB2GW7. The recombinant construct was transferred into Agrobacterium tumefaciens strain GV3101, and was used for Agrobacterium mediated transformation of tobacco plants. The presence of the desired gene in transgenic lines was confirmed through colony PCR. The expression of the protein in transgenic lines was confirmed by immune-dot blot assay and ELISA. Although the transformation of the two species was confirmed by immune-dot blot assay and SDS-PAGE, recombinant protein production in Nicotiana tabacum plants was confirmed by ELISA and it was estimated 400 µg per gram wet weight of tobacco leaves. According to the results, this species is the appropriate host for the production of recombinant HDA19, one of the histone deacetylases, rather than Nicotiana bentamiana.

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References

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