PaADA
(Plasmid #250880)

• Purpose: Pseudomonas aeruginosa adenine deaminase (PaADA; Pa0148; UniProt Q9I6Y4)
• Depositing Lab: Vern Schramm
• Publication: Coupled-Enzyme Assay for MTAP Activity in Biological Samples (unpublished)

Backbone

• Vector backbone: pET-28a(+)
• Backbone manufacturer: Genscript
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: The construct was transformed into E. coli One Shot™ BL21(DE3) pLysS cells (Thermo Fisher Scientific, Cat. #C606010). Single colonies were inoculated into 20 mL LB medium supplemented with 50 µg/mL kanamycin and 30 µg/mL chloramphenicol and cultured overnight at 37 °C. The resulting pre-cultures were used to inoculate 1 L LB medium containing the same antibiotics, grown at 37 °C to an optical density of 0.5 at 600 nm before induction with 1 mM IPTG. After 20 h at 20 °C, cells were collected by centrifugation. Protein purified by Ni-NTA. TEV cleavable 6xHis Tag.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Pseudomonas aeruginosa adenine deaminase (PaADA; UniProt Q9I6Y4)
• Alt name: Pa0148
• Entrez Gene: PA0148 (a.k.a. PA0148)
• Tag / Fusion Protein:
  • 6xHis-TEV- (N terminal on insert)

Resource Information

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

Terms and Licenses

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