1a.Objectives (from AD-416):
Molecular Analysis of Pecan Nut Development.
1b.Approach (from AD-416):
Samples of developing Pecan nuts will be extracted to isolate total ribonucleic acid (RNA) and protein. RNA will be sequenced using next-generation sequencing, and then analyzed and annotated to identify specific genes and compare expression profiles. Quantitative-real-time polymerase chain reaction (qRT-PCR) will be done to confirm findings for individual genes. Total protein accumulation during Pecan nut development will be analyzed by gel electrophoresis and mass-spectrometry. Major Pecan allergens Car i 1, Car i 4, and the 7S vicllin will be studied using each of these techniques to characterize their expression, processing, and accumulation. Other Pecan allergens will also be identified based upon homology.
Funds for this project have just recently been made available. Supplies are being purchased for the project, samples are being collected, and we are evaluating sample processing methods to optimize isolated Rubric Nucleic Acid (RNA) and protein quality for downstream steps of the study.