hSDH5
(Plasmid #113046)

• Purpose: Expression of WT hSDH5
• Depositing Lab: Edward A. Berry
• Publication: Zafreen et al Arch Biochem Biophys. 2016 Aug 15;604:47-56. doi: 10.1016/j.abb.2016.06.006. Epub 2016 Jun 11.

Backbone

• Vector backbone: pET41
• Modifications to backbone: For expression of hSDH5 in E. coli, we designed a putative mature sequence starting with 39SPTD. Primers were designed to amplify this sequence with extensions to insert it between the NdeI and XhoI restriction sites of the pET41 vector. The bases of the NdeI site provide the initiator Met while the XhoI site and following bases of the vector append LEH8 on the C terminus. A second construct, in which nucleotides coding for the His8 tag were deleted but the linker residues LE encoded by the restriction site were retained, was designed for expression of tag-less mature hSDH5 for pulldown studies with H6hSDHA. and conventional purification. It is this tag-less, mature construct hSDH5, that is being deposited.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): BL-21
• Growth instructions: For protein expression (induced with IPTG) grow at 30C but for growing cells without IPTG grow at 37C.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: hSDH5
• Alt name: SDHAF2
• Species: H. sapiens (human)
• Insert Size (bp): 393
• Mutation: Insert begins at amino acid 39
• GenBank ID: JF432739.1
• Entrez Gene: SDHAF2 (a.k.a. C11orf79, PGL2, SDH5)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: unknown (unknown if destroyed)
• 3′ cloning site: unknown (unknown if destroyed)
• 5′ sequencing primer: unknown

Terms and Licenses

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