pET28a MDHP_YEAST
(Plasmid #204233)

• Purpose: Bacterial expression of yeast peroxisomal malate dehydrogenase ; Use in CURES (integration of research into undergraduate teaching labs).
• Depositing Lab: Malate dehydrogenase CUREs Community (MCC)
• Publication: Expression of His tagged recombinant malate dehydrogenase (MDH) (unpublished)

Backbone

• Vector backbone: pET28a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5235
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: MDHP_YEAST
• Alt name: Yeast peroxisomal MDH
• Alt name: peroxisomal yeast MDH, Yeast MDH3
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 1068
• Mutation: The last 3 C-terminal residues (SKL) are the peroxisomal transit signal and were left in the final version.
• GenBank ID: NM_001180137.1
• Entrez Gene: MDH3 (a.k.a. YDL078C)
• Promoter: Lac
• Tag / Fusion Protein:
  • 6X His (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NcoI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: T7 Promoter
• 3′ sequencing primer: T7 Terminal

Resource Information

• Supplemental Documents:
  • pET28a-MDHP_YEAST SnapGene
  • MCC Clone Information Sheet - MDHP_YEAST

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

UniProt ID: https://www.uniprot.org/uniprotkb/P32419/.

The affinity of MDH3 for OAA is lower than those of MDH1 and MDH2. The structure of the ternary complex suggests that the active-site loop is in the open conformation, differing from the closed conformations in mitochondrial and cytosolic malate dehydrogenases.

For more information, please see the MDH CUREs Community website: https://www.mdhcures.org/.