pET28a MDHM_YEAST
(Plasmid #204231)

• Purpose: Bacterial expression of yeast mitochondrial 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• Publication: Expression of His tagged recombinant malate dehydrogenase (MDH) (unpublished)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): 5320
• 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: MDHM_YEAST
• Alt name: Yeast mitochondrial MDH
• Alt name: Yeast MDH1, ymMDH
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 993
• Mutation: Does not include transit peptide and includes a Met for coding
• GenBank ID: NM_001179651.1
• Entrez Gene: MDH1 (a.k.a. YKL085W)
• 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 MDHM_YEAST SnapGene
  • MCC Clone Information Sheet - MDHM_YEAST

Terms and Licenses

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

Depositor Comments

UniProt ID: https://www.uniprot.org/uniprotkb/P17505/.

Expression changes based on metabolite starting point. Involvement of MDH in the asp-malate shuttle in yeast extends replicative lifespan and may play a role in calorie-restricted life extension.

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