Two-dimensional electrophoresis of acidic proteins isolated from ozone-stressed Norway spruce needles (Picea abies L. Karst): Separation method and image processing
✍ Scribed by Dr. Nina B. Davidsen
- Publisher
- John Wiley and Sons
- Year
- 1995
- Tongue
- English
- Weight
- 613 KB
- Volume
- 16
- Category
- Article
- ISSN
- 0173-0835
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✦ Synopsis
Pwo-dimensional electrophoresis of acidic proteins isolated from ozone-stressed Norway spruce needles processing Forest Research Institute, (Picea abies L. Karst): Separation method and image
The effect of ozone stress (< 10,200 or 400 Vg m-3) on the protein pattern of Picea abies L. Karst needles was studied by fumigating potted grafts from mature trees. Clonal trees of Norway spruce were fumigated in open-top chambers for one growing season. Proteins, soluble and membrane bound, were extracted from the needles in a four-step procedure: (i) grinding in liquid nitrogen and then in a pH 5.0 buffer with thiourea added to inhibit phenol oxidase and polyvinylpolypyrrolidone (PVP) added to bind phenolic compounds; (ii) precipitation of proteins with acetone at -20°C; (iii) solubilization in a modified lysis buffer; and (iv) dialysis against the modified lysis buffer. Proteins were separated by two-dimensional electrophoresis according to O'Farrell's method. The changes in the silver-stained protein pattern were evaluated by using image processing methods. Electrophoretograms from stressed and nonstressed trees are presented.