expression of a taste-modifying protein, miraculin, in by-2 tobacco cell line (Nicotiana tabacum L. Cv Bright Yellow 2)

Authors

  • La Viet Hong VAST
  • Nguyen Thu Giang
  • Pham Bich Ngoc
  • Chu Hoang Ha

DOI:

https://doi.org/10.15625/0866-7160/v36n3.5977

Keywords:

Arabidopsis, Nicotiana, Richadella, BY-2, diabetes, miracle fruit, miraculin, taste-modifying protein

Abstract

Miraculin is a taste-modifying and low calorie protein isolated from the red berries of Richadella dulcifica, a shrub native to West Africa. In this study, a synthetic miraculin gene was designed to optimize its codon usage for BY-2 tobacco cell lines. This gene was inserted into two constructs including pBI121/HSP-pro:Mir:HSP-ter that contains HSP promoter and HSP terminator isolated from Arabidopsis thaliana for enhancing production of miraculin and pBI121/35S-pro:Mir:NOS-ter for transformation into BY-2 tobacco cell lines. The presence and expression of miraculin gene in BY-2 tobacco cell lines were verified by PCR and Western blot, respectively. It was demonstrated that transgenic BY-2 tobacco cell lines grew stably in liquid media after 4-5 times of transplant. The expression of miraculin gene in transgenic BY-2 tobacco cell lines resulted in the accumulation of miraculin protein with molecular mass of approximately 43-45 kDa. Besides, it was observed that the expression of miraculin in HSP-pro:Mir:HSP-ter transformed BY-2 tobacco cell lines is better than the expression of miraculin in 35S-pro:Mir:HSP-ter transformed ones. For the first time, miraculin recombinant protein has been expressed in BY-2 tobacco cells providing a great potential for research and production of miraculin protein at industrial scale.

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Published

19-03-2015

How to Cite

Hong, L. V., Giang, N. T., Ngoc, P. B., & Ha, C. H. (2015). expression of a taste-modifying protein, miraculin, in by-2 tobacco cell line (Nicotiana tabacum L. Cv Bright Yellow 2). Academia Journal of Biology, 36(3), 367–372. https://doi.org/10.15625/0866-7160/v36n3.5977

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