pALOD4 non-binding mutant
(Plasmid #111027)

• Purpose: Expresses non-binding mutant version of p-ALOD4, has six inactivating mutations (G501A, T502A, T503A, L504A, Y505A, and P506A)
• Depositing Lab: Arun Radhakrishnan
• Publication: Gay et al Biophys J. 2015 Mar 24;108(6):1459-69. doi: 10.1016/j.bpj.2015.02.008.

Backbone

• Vector backbone: pRSET-B
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 2887
• Modifications to backbone: None
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: BL21 (DE3) pLysS E. coli-competent cells used for protein production. Growth at 37°C until O.D. of 0.6 - 0.8. Then, expression induced with 1 mM IPTG at 18°C for 16 h
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Domain 4 of Anthrolysin O
• Alt name: ALOD4
• Species: Bacillus Anthracis
• Insert Size (bp): 429
• Mutation: Mutations: S404C, C472A, G501A, T502A, T503A, L504A, Y505A, P506A
• GenBank ID: AAT_32463.1
• Promoter: T7
• Tag / Fusion Protein:
  • His6 tag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamH1 (unknown if destroyed)
• 3′ cloning site: EcoR1 (unknown if destroyed)
• 5′ sequencing primer: TAA TAC GAC TCA CTA TAG GG
• 3′ sequencing primer: GCT AGT TAT TGC TCA GCG G

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Genscript, Inc.

Terms and Licenses

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