pET24a MDH_HALMA
(Plasmid #204241)

• Purpose: Bacterial expression of H. marismortui (Dead Sea halophilic red archaeon) cytosolic 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• Publication: Expression of His tagged recombinant malate dehydrogenase (MDH) (unpublished)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: pET24a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5238
• 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: MDH_HALMA
• Species: Haloarcula marismortui
• Insert Size (bp): 950
• GenBank ID: NZ_CP039138.1
• Entrez Gene: mdh (a.k.a. E6P14_RS15400, E6P14_15395)
• 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:
  • pET24a MDH HALMA SnapGene
  • MCC Clone Information Sheet - MDH_HALMA

Terms and Licenses

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

Depositor Comments

UniProt ID: https://www.uniprot.org/uniprotkb/Q07841/.

Chloride ions bound between subunits stabilize the quaternary structure. This may be an adaptation to the halophilic environment. Sequence is similar to LDH, more than other MDH isoforms. Generates an active tetramer upon ~3-4 M NaCl.

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