Abstract
Nicotiana transformed with the isopentenyl transferase (ipt) gene from Agrobacterium tumefaciens was fixed for 1 h in 1% glutaraldehyde and 4% formaldehyde. Ultrathin sections were collected on nickel grids. Grids were treated with polyclonal anti-IPTase antibody raised in rabbits and visualized with 10 nm, protein-A–labeled colloidal gold. Gold label was found throughout the cell, including the cell wall, vacuole, rough ER, and organelles. Cell wall and vacuole labeling appears to be due to non-specific binding and is greatly reduced by a BSA block. Mitochondria and chloroplasts also showed gold label, but not greater than established background levels. Labeling above background levels on the rough ER, free polysomes, and further label in the free cytoplasm indicate a cytoplasmic role for IPTase.
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