Team:Valencia UPV/Project/modules/biosafety

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Biosafety

In order to make the Sexy Plant a safe plant, we developed a biosafety module which will be available for teams working with plants in future iGEM editions. The aim of the biosafety module is to avoid the dispersion of the plant’s genetic material in the environment. Two strategies were combined into the same module to make our plant safe: identity preservation and male sterility.



Identity preservation enables an easy identification of the genetically modified organism. A chromoprotein, which provides a differential pigmentation to the plant, is incorporated in the module. The implementation of this chromoprotein to our system is the result of the collaboration with NRP-UEA-Norwich, and it is also used in their project “Green Canary”.



Male sterility makes impossible the dispersion of genetic material using pollen as the vehicle. In order to achieve this dispersion restriction, the gene coding for barnase (a ribonuclease from Bacillus amyloliquefaciens) under the regulation of the tapetum-specific promoter TA29[1] is incorporated in our system. Both components are very well documented since TA29 has been used by a large number of researchers [2-6] and barnase has also been used under the regulation of different promoters [7,8]. This strategy was chosen because it had been previously used in our laboratory with satisfactory results [9]



References


  1. Mariani C, Beuckeleer MD, Truettner J, Leemans J, Goldberg RB (1990) Induction of male sterility in plants by a chimaeric ribonuclease gene. Nature 347: 737-741.
  2. Cho HJ, Kim S, Kim M, Kim BD (2001) Production of transgenic male sterile tobacco plants with the cDNA encoding a ribosome inactivating protein in Dianthus sinensis L. Mol Cells 11: 326-333.
  3. Sa G, Mi M, He-Chun Y, Guo-Feng L (2002) Anther-specific expression of ipt gene in transgenic tobacco and its effect on plant development. Transgenic Res 11: 269-278.
  4. Shaya F, Gaiduk S, Keren I, Shevtsov S, Zemah H, et al. (2012) Expression of mitochondrial gene fragments within the tapetum induce male sterility by limiting the biogenesis of the respiratory machinery in transgenic tobacco. J Integr Plant Biol 54: 115-130.
  5. Kriete G, Niehaus K, Perlick AM, Puhler A, Broer I (1996) Male sterility in transgenic tobacco plants induced by tapetum-specific deacetylation of the externally applied non-toxic compound N-acetyl-L-phosphinothricin. Plant J 9: 809-818.
  6. Shukla P, Singh NK, Kumar D, Vijayan S, Ahmed I, et al. (2014) Expression of a pathogen-induced cysteine protease (AdCP) in tapetum results in male sterility in transgenic tobacco. Funct Integr Genomics 14: 307-317.
  7. Goldman MH, Goldberg RB, Mariani C (1994) Female sterile tobacco plants are produced by stigma-specific cell ablation. EMBO J 13: 2976-2984.
  8. Wang HZ, Hu B, Chen GP, Shi NN, Zhao Y, et al. (2008) Application of Arabidopsis AGAMOUS second intron for the engineered ablation of flower development in transgenic tobacco. Plant Cell Rep 27: 251-259.
  9. Sarrion-Perdigones A, Falconi EE, Zandalinas SI, Juarez P, Fernandez-del-Carmen A, et al. (2011) GoldenBraid: an iterative cloning system for standardized assembly of reusable genetic modules. PLoS One 6: e21622.
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