Solution requests schedule

Project Literature

Gene Cloning

Cloning of yeast FUM1 gene by Wu and Tzagoloff

Protein Purification

Salting out yeast fumarase with ammonium sulfate
A new affinity purification of porcine fumarase
Purication of fumarase(s) from yeast
Purification and crystalization of E. coli fumarase by Weaver et al.
Purification, characterization and preliminary X-ray study of Fum1p

Protein Structure

Mutagenic distinction of active and second site of E. coli fumarase by Weaver et al.
Crystallography of active and second site of E. coli fumarase by Weaver et al.
Fumarase active site modeling by Rose
Structure of native and recombinant yeast fumarase
Analysis of a human fumarase mutation

Cellular Distribution and Mitochondria Import

Distinction of cytosolic and mitochondrial fumarases
See also Wu and Tzagoloff in Gene Cloning
Inducible overexpression of Fum1p and localization by Peleg et al.
Mitochondrial processing of Fum1p and cellular distribution by Stein et al.
Cellular distribution of yeast fumarase by Wu et al.
Mitochondrial import of Fum1p by Knox et al.
Cellular distrubution of yeast Fum1p by Sass et al.

Other

Fumarase formation in mitochondria biogenesis
Genetic control of metabolic modifications during sporulation
Fumarate overproduction in fum1 mutants
Cytosolic L-malate production by Pines et al.
Transcriptional control of TCA genes and respiration
Interactions of TCA cycle gene products
Association of fumarase and malate dehydrogenase

Novagen HisBind Resin
YeastBuster Protein Extraction Reagent
Chemicon mouse anti-actin antibody
US Biolgical rabbit anti-histidinyl antiserum

CHE 255 Laboratory Manual
Amersham site on comparison of metals in IMAC
Information on the Sims-Aminco French press (use Internet Explorer)
Handling and care of a French pressure cell
Use of the French pressure cell
Information on the BioSpec Products BeadBeater^TM
Review on Cell Disruption at BioSpec
Second review on Cell Disruption
Yeast cell lysis with a French press
General tips on cell lysis with a French press
Instructions for BioRad slot-blot apparatus

W303DFUM1 genotype - MATa ade2-1 his3-11,15 leu2-3,112 trp1-1 ura3-1 fum1::LEU2
Maintenance and culture of baker's yeast

Solution maker schedules
    Monday 10:30-12:30
    Tuesday 2:30-4:30
    Thursday 10:30-12:30
    Friday 10:30-12:30

synthetic yeast media lacking uracil

YEP-GAL and galactose solutions, YEP-dextrose

YEP-Lactate
    Mix Carbon solution and Media solution 1:1 for use.

    Carbon solution
        2% (w/v) lactate (using the pentahydrate calcium salt, 34.6 grams/liter)
        adjust pH to 5.5
        autoclave to sterilize
    Media solution
        2% (w/v) yeast extract
        4% (w/v) peptone
        0.1% (w/v) dextrose
        0.006% (w/v) adenine sulfate
        adjust pH to 5.5
        autoclave to sterilize