Expression of interleukin-4delta2 in plants

Abstract : There is an increasing demand for protein-based therapeutics. Unlike small molecules, which can be synthesized by chemical means, most proteins necessitate their production in recombinant living systems such as animal cells, yeast and bacteria. These past decades have seen the emergence of plants as an alternative protein production platform. In this study, the overall aim was to develop a novel recombinant protein that could boost the protective immune response in individuals latently infected with Mycobacterium tuberculosis. The target protein is a spliced form of the cytokine interleukin-4 called interleukin-4delta2 (IL-4delta2), and the objective of the project was to assess the feasibility of expressing this cytokine in Nicotiana benthamiana plants. The cDNA of IL-4delta2 was cloned and inserted into two different plant expression vectors. These vectors were then used for transient expression in Nicotiana benthamiana, using Agrobacterium tumefaciens-mediated delivery. Expression of IL-4delta2 was confirmed by western blot. The plant extracts contained the recombinant IL-4delta2 protein to approximately 0.002% of total soluble protein. Expression of two control recombinant proteins confirmed that the expression system was functional and that the low protein expression of IL-4delta2 was gene specific. The low protein expression yield was investigated further and a mutated version of the gene of interest was proposed, engineered and cloned.
Document type :
Master thesis
File URL :
http://docnum.univ-lorraine.fr/prive/SCDSCI_M_2011_MORVAN_LIONEL.pdf
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https://hal.univ-lorraine.fr/hal-01788677
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Submitted on : Wednesday, May 9, 2018 - 12:16:51 PM
Last modification on : Friday, September 14, 2018 - 1:01:46 AM

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  • HAL Id : hal-01788677, version 1

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Lionel Morvan. Expression of interleukin-4delta2 in plants. Sciences de l'environnement. 2011. ⟨hal-01788677⟩

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