| Products/Services Used | Details | Operation |
|---|---|---|
| Peptide Synthesis | For western blot analysis, concentrated PGIP protein extracts (20 µg total) were electrophoresed on 12% Tris-Glycine mini gels (XCell SureLock Mini-Cell, Invitrogen, Carlsbad, CA). Separated proteins were transferred to polyvinylidene difluoride (PVDF) membranes using the XCell II Blot Module apparatus (Invitrogen) for 1 h 30 min. Membrane blotting and detection of primary antibody were carried out using the WesternBreeze Chromogenic Immunodetection Kit (Invitrogen). Briefly, transferred membranes were incubated with rabbit polyclonal antibody against BvPGIP1 peptide (CVQITEAQDIYGPIP) (GenScript Corporation, Piscataway, NJ) at 1:2000 dilution for 1 h and then incubated for 30 min with alkaline phosphatase conjugated secondary antibody (Goat Anti-Rabbit IgG). Alkaline phosphatase was detected with Chromogenic NBT/BCIP Substrate until purple bands developed on the membrane. The relative levels of the accumulated BvPGIP proteins were estimated by densitometry with an AlphaImager HP. | Get A Quote |
Soil-borne fungus Fusarium oxysporum f. sp. betae (Fob) is the causative agent of Fusarium yellows in sugar beet. Leaf interveinal yellowing and root vascular discoloration significantly reduce root yield as well as sucrose content and juice purity. Fob, like other fungal pathogens, initiates disease development by secreting polygalacturonase (PG) enzymes to break down plant cell walls during early stages of infection. To protect themselves, plants produce polygalacturonase-inhibiting proteins (PGIPs). In our study of sugar beet root defense responses, several PGIP genes (BvPGIPs) were identified. To determine if BvPGIPs inhibit Fob PGs, genes BvPGIP1, BvPGIP2 and Bv(FC607)PGIP1 were fused with the C... More
