Abstract: The embodiments herein provide methods to analyze the in vitro stability of recalcitrant or membrane-bound proteins in simulated gastric fluid (SGF) comprising the proteolytic enzyme, pepsin, and in combination with a novel pepsin-trypsin assay employing state-of-the-art mass spectrometric approaches, such as LC-MS/MS, to monitor the precise degradation products. The extent of protein digestion can be evaluated by the appearance of peptic products and the disappearance of tryptic peptide products (as a proxy for intact protein). The embodiments herein also provide methods for protein quantitation using high-sensitivity LC-MRM-MS quantification. The methods embodied herein are particularly useful in charactering proteins produced in transgenic plants, such as canola genetically engineered to produce long chain omega-3 polyunsaturated fatty acids.

Abstract: The present invention relates to cyclotides and cyclotide-encoding genes from the Fabaceae plant family, and to the expression of cyclotides in Fabaceae. The present invention further relates to isolated nucleic acids configured to express cyclotides comprising heterologous peptide grafts in plants of the Fabaceae family.

Abstract: The present invention relates to cyclotides and cyclotide-encoding genes from the Fabaceae plant family, and to the expression of cyclotides in Fabaceae. The present invention further relates to isolated nucleic acids configured to express cyclotides comprising heterologous peptide grafts in plants of the Fabaceae family.