To generate transgenic soybean lines, coding sequences of the human growth hormone; human coagulation factor IX, NY-ESO-1 and cyanovirin CV-N genes were placed under control of different seed-specific promoters and signal peptides. Utilizing biolistic-mediated transformation Rech et al.
Recombinant Proteins From Plants - Methods and Protocols | Loïc Faye | Springer
Accumulation in leaf tissue was influenced by the subcellular localization. No signal was detected when the NY-ESO-1 antigen was targeted to chloroplast or to the apoplast, and detection was only possible when the protein was targeted to the ER. Exploiting the utilization of seed-specific regulatory sequences was possible to direct and accumulate recombinant proteins in the PSV's from soybean seeds [ 5 ].
The somatogenic activity bioassay demonstrated that the hGH expressed in soybean seeds is fully active. Protein extracts from transgenic seeds containing the hFIX showed a blood-clotting activity of up to 1. Purification of pure recombinant CV-N in soybean seeds is still ongoing, showing a high affinity for CV-N to bind to glycinin and B-conglycinin proteins from seeds.
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NanoUPLC-MS e analytical results indicated expression level and correct structural characterization of recombinant protein sequences. Ultrastructural immunocytochemistry assays revealed that the recombinant proteins accumulated in the endoplasmic reticulum ER -derived protein storage vacuoles. Recombinant molecules were visualized all along the cistern lumen but were absent from the interior of seed oil bodies.
This should contribute to: a understanding the minimum number of bioparts necessary for metabolic function and regulation of gene expression; b developing synthetic chromosomes, which may enable the accurate expression of gene families; c developing regulatory circuits and designing synthetic genes that can be based on standard biological parts, as models for the development of synthetic biology tools for engineering genomes.
In addition, making use of genomics as a template founded on molecular synthesis and assembly, using recombinant DNA technology and synthetic biology, has allowed the sustainable prospection and manipulation of innovative traits found in biodiversity [ 10 ]. The aim includes improvement and development of novel tools for production of recombinant proteins in plant systems.
Recombinant Proteins from Plants
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