Purification of vacuoles from Neurospora crassa.

@article{Vaughn1981PurificationOV,
  title={Purification of vacuoles from Neurospora crassa.},
  author={L. E. Vaughn and Rowland H. Davis},
  journal={Molecular and cellular biology},
  year={1981},
  volume={1 9},
  pages={797-806}
}
The Neurospora crassa vacuole, defined by its content of basic amino acids, polyphosphate, protease, phosphatases, and alpha-mannosidase, was purified to near homogeneity. The procedure depends upon homogenization of snail gut enzyme-digested cells in a buffer osmotically stabilized with 1 M sorbitol, differential centrifugation of the extract, and sucrose density gradient centrifugation of the organellar pellet. Isopycnic centrifugation of vacuoles in 2.25 M sorbitol-Metrizamide density… CONTINUE READING