pBtPIPLC His
(Plasmid #173808)

• Purpose: Expression and purification of the mature form of Bacillus thuringiensis phosphaatidyl inositol specific phospholipase C (PI-PLC), UniProt ID P08954 from E. coli BL21 CodonPlus (DE3) RIL cells
• Depositing Labs: Anne Gershenson, Mary F Roberts
• Publication: He et al J Biol Chem. 2015 Jul 31;290(31):19334-42. doi: 10.1074/jbc.M115.668343. Epub 2015 Jun 19.

Backbone

• Vector backbone: pET-23b
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: E. coli BL21-Codonplus (DE3)-RIL can be used for expression. Note that this protein purification is performed at room temperature.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: Bacillus thuringiensis 1-phosphatidylinositol phosphodiesterase
• Alt name: Bacillus thuringiensis phosphatidyl-inositol specific phospholipase C
• Alt name: BtPI-PLC
• Species: Other
• Mutation: none
• Promoter: T7
• Tag / Fusion Protein:
  • 6XHis (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Unknown (unknown if destroyed)

Terms and Licenses

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

Depositor Comments

Bacillus thuringiensis PI-PLC (UniProt P08954) cleaves GPI-anchored proteins off the surface of cells and can be used instead of the 97% identical, commercially available B. cereus enzyme (UniProt P14262).

Also see PubMed 30148347 (bacterial PI-PLC review).