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DESIGN strategies to produce high levels of recombinant proteins and peptides in plants as bioreactors according to an in silico pipeline

The known structural features of the protein of interest (POI) are used for in silico modelling of the POI, and the choices of vector, target sequence, codon-usage, expression system and purification strategy are tailored to the required POI properties (e.g. stability, purity, or alleviation of allergenic/toxic features).

TESTING the different strategies by ...

- In silico modelling of recombinant proteins for optimization of their production in plants, including post-translational modifications and prediction of subcellular localisationAnalysis of:

- Folding: resistance to proteolytic digestion, disulfide bond pattern, solubility, binding to conformation-dependent antibodies.

- Oligomerization: velocity gradient centrifugation in different biochemical conditions; dynamic light scattering (DLS); SEC-MALS.

- Glycosylation: analysis with glycan-specific ...

Based on the following:

-       Extraction of genomic DNAs directly from the environmental samples;

-       Preparation of metagenomic libraries;

-       Shotgun metagenomic sequencing by Next generation sequencing using Illumina technology (Metagenomics);

-       Bioinformatics analyses (meta-assembly, gene findings and annotation, metabolic reconstruction) and data delivery;

-       16S rRNA and other target genes sequencing by Next generation sequencing using Illumina technology (Metabarcoding) ...

Structure determination using experimental and computational resources (virtual machine), based on X-ray diffraction and/or Cryo-electron microscopy.