Project description:Purpose: Analyses of transcriptomes of WT and pumpkin mutants, in order to compare gene expression of typically NEP or PEP-transcribed plastid genes.
Project description:Stroma extracts were isolated from 2-week-old WT plants and incubated with either PUMPKIN-specific antibodies or with the pre-immune serum. IgGs were captured with SiMAG-Protein G beads (Chemicell) and recovered RNA was used for generation of libraries with the ScriptSeq v2 RNA-seq Library Preparation Kit (Epicentre). Primary reads were aligned to the Arabidopsis chloroplast genome (accession number NC_000932.1) using CLC Genomics Workbench, the mean RPKM values of the replicates as well as the ratio of PUMPKIN vs Pre-immuneserum were calculated. Five prominent RNA targets (trnG-UCC, trnV-UAC, petB, petD and ndhA) were identified and validated via Slot Blot analyses.
Project description:Food fraud is a common issue in the modern food industry. The undeclared use of foreign pro-teins in meat products is a major concern in this context. Oilseeds are ideal for this purpose due to their high protein content and since huge amounts of oil meals are obtained as a by-product of oil production. Therefore, a UHPLC-MS/MS method was developed for the simultaneous de-tection of chia, coconut, flaxseed, hemp, peanut, pumpkin, rapeseed, sesame, soy, and sunflower proteins in meat products. Potential tryptic peptide markers were identified by high-resolution mass spectrometry. The final twenty peptide markers selected, which are specific for one of the ten species targeted were each measured by multiple reaction monitoring. To the best of our knowledge, twelve new heat-stable marker peptides for chia, coconut, flaxseed, pumpkin, rape-seed, sesame and sunflower have not been reported previously.