Disruption of fungal cells

[Michel Traris]

Hi,
 I would like to disrupt the cells of the filamentous fungi METHARHIZIUM in
order to extract cytosolic proteins. I tried already the following procedur=
e:

1. Filter the fungal culture=20
2. Freeze wet mycelium with liquid nitrogen.=20
3. Grind rigorously in a pre-cooled mortar with stepwise addition of buffer
(with protease inhibitor) and liquid nitrogen=20
4. Stir the homogenate in a baker with one volume of glass beads (1 mm) at =
4=A1C.

The result is not satisfying as I can see a lot of long unbroken hyphae und=
er
the microscope. Any protocols or suggestion for an improvement of the
extraction procedure would be highly welcomed.

Thank you=20
Michel.

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Michel TRARIS
Laboratoire de Mycologie
Station de Pathologie Comparee INRA-CNRS
30380 St-Christol-lez-Ales=09tel:66 78 37 00=09=09fax:66 52 46 99
France                          E-mail:traris at ales.inra.fr
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