pOlyA(WT)
(Plasmid #132391)

• Purpose: Ostreolysin A for sensing sphingomyelin/cholesterol complexes
• Depositing Lab: Arun Radhakrishnan
• Publication: Endapally et al Cell. 2019 Feb 21;176(5):1040-1053.e17. doi: 10.1016/j.cell.2018.12.042. Epub 2019 Jan 31.

Backbone

• Vector backbone: pET21c+
• 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): NEB Stable
• Growth instructions: BL21 (DE3) pLysS E. coli-competent cells used for protein production. Growth at 37°C until O.D. of 0.4 - 0.8. Then, expression induced with 1 mM IPTG at 18°C for 16 h
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Ostreolysin A wild-type
• Alt name: OlyA(WT)
• Species: Pleurotus ostreatus
• Insert Size (bp): 480
• Mutation: Mutations: C62S, C94S, S151C
• GenBank ID:
• Tag / Fusion Protein:
  • His6 tag (C terminal on insert)

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Kristina Sepcic, University of Ljubljana, Ljubljana, Slovenia

Terms and Licenses

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

Depositor Comments

Additional Reference:
Skocaj M, Resnik N, Grundner M, Ota K, Rojko N, et al. (2014) Tracking Cholesterol/Sphingomyelin-Rich Membrane Domains with the Ostreolysin A-mCherry Protein. PLoS ONE 9(3): e92783. doi:10.1371/journal.pone.0092783