pT22-GluCNZ-6His
(Plasmid #213504)

• Purpose: Designed for cytoplasmic E.coli overexpression of glucanase (β-1,3-glycosidase) for gentle yeast spheroblasting that unlike Zymolyase prevents proteolysis of intracellular components.
• Depositing Lab: John Denu
• Publication: Kuznetsov et al J Biol Chem. 2018 Aug 3;293(31):12271-12282. doi: 10.1074/jbc.RA118.002984. Epub 2018 May 24.

Backbone

• Vector backbone: pet22B
• Backbone manufacturer: Novagen
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: Glucanase (glucan endo-1,3-beta-glucosidase)
• Alt name: Lyticase
• Alt name: Zymolyase
• Species: Cellulosimicrobium cellulans (Arthrobacter luteus)
• Insert Size (bp): 1580
• Mutation: 1-36 Signal Sequence Deletion
• Promoter: T7
• Tag / Fusion Protein:
  • 6His tag (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7-promotor
• 3′ sequencing primer: T7-terminator

Terms and Licenses

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