pMBP-hLIF
(Plasmid #179711)

• Purpose: Expresses H6-MBP-LIF
• Depositing Lab: Mark Arbing
• Publication: Bacterial expression of Leukemia inhibitory factor as a maltose binding protein fusion protein (unpublished)

Backbone

• Vector backbone: pMAL-C2
• Backbone manufacturer: New England Biolabs
• Backbone size w/o insert (bp): 6600
• Total vector size (bp): 7162
• Modifications to backbone: Add N-terminal His6 tag to MBP.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Leukemia inhibitory factor
• Alt name: LIF
• Species: H. sapiens (human)
• Insert Size (bp): 543
• GenBank ID: M63420
• Entrez Gene: LIF (a.k.a. CDF, DIA, HILDA, MLPLI)
• Promoter: tac
• Tag / Fusion Protein:
  • His6-MBP-TEV protease site (N terminal on backbone)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: M13 reverse
• 3′ sequencing primer: M13 forward

Resource Information

• Supplemental Documents:
  • pmal-h6-mbp-hlif.gb

Terms and Licenses

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

Depositor Comments

Express fusion protein in E. coli BL21-Gold DE3 at 18C overnight. Lysis buffer: 20 mM Tris pH 8, 300 mM NaCl, 10% glycerol. Purification by NiNTA and size exclusion (Superdex 200) chromatography. LIF precipitates when cleaved from MBP however the fusion protein is functional.