Objective
The objective of this proposal is to use plants for producing bovine FSH and its receptor, and to apply these products for the purpose of assisted reproduction in cattle. This will be done by constructing vectors of Tobacco Mosaic Virus, which will allow separate expression of foreign proteins through a subgenomic promoter.
In order to allow for high biological activity in-vivo, plant glycosylation will first be tailored towards a more mammalian type, with terminal sialic acid attached to glycosidic sidechains. Purified plant gonodotrophin will be assessed for its use as therapeutic agent to improve fertility in cows.
Corresponding purified receptors will be used to develop a test kit for diagnosis of fertility problems, and for monitoring of treatment effects.
The proposal involves:
1. preparation of cDNA of receptor for bovine FSH
2. construction of TMV vectors carrying genes of bovine FSH and its corresponding receptor
3. construction of transgenic plants expressing beta 1,4 galactosyltransferase and alpha 2,6 sialyltransferase, and construction of transgenic plants that lack immunogenic alpha 1,3 fucose and beta 1,2 xylose
4. production of plant-secreted recombinant bovine FSH and its corresponding receptor
5. construction of transfected cell-lines expressing bovine FSH-receptor, and development of an homologous in-vitro bioassay
6. downstream processing and upscaling of production of FSH and receptor
7. development and validation of receptor assay for bovine FSH
8. clinical trials to assess tolerance, pharmacokinetics and therapeutic efficacy of plant-produced recombinant bovine FSH
Plant-secreted recombinant gonadotrophin will improve biosafety by eliminating the risk of contamination with prior-like proteins, which can cause BSE-related disease, or with unwanted hormones, which can cause ovulation of premature eggs. Development of in-vitro bioassays for the measurement of gonadotrophins will reduce the need for experimental animals in in-vivo bioassays. Whole plants have the advantage over plant cells in that there is no need for sterile conditions, and they are much easier and cheaper to grow. It is not the plant, however, but the virus that is used as the recombinant vehicle. Production will take place in specially equipped laboratories and insulated greenhouses. This project will show the way to use plants as bioreactors for production of complex bioactive proteins.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques.
- natural sciencesbiological sciencesmicrobiologyvirology
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteins
- social sciencessociologydemographyfertility
- medical and health sciencesbasic medicinepharmacology and pharmacypharmacokinetics
- agricultural sciencesanimal and dairy sciencedomestic animalsanimal husbandry
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Topic(s)
Call for proposal
Data not availableFunding Scheme
CSC - Cost-sharing contractsCoordinator
8200 AB Lelystad
Netherlands