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pET24a_UDP_Xase_E162
(Plasmid #73160)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 73160 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pET24
  • Backbone manufacturer
    Novagen
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    For protein production, use GC5 cells.
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    pET24a
  • Alt name
    A0A0H2URA0
  • Alt name
    NC_003028.3
  • Alt name
    Sp_1699
  • Species
    Streptococcus pneumoniae serotype 4 (strain ATCC BAA-334 / TIGR4
  • Mutation
    change glutamate 162 to alanine
  • GenBank ID
    NC_003028.3

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BAMH1 (not destroyed)
  • 3′ cloning site NdeI (not destroyed)
  • 5′ sequencing primer 5 9 -TGGACAATTCCAA- GAAAATCAAGCAATTGCTGACCTTCAATTTTTTG-3
  • 3′ sequencing primer 5 9 -CAAAAAATTGAAGGTCAGCAATTGCTT- GATTTTCTTGGAATTGTCCA-3 9 ,
    • (Common Sequencing Primers)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Cells expressing proteins containing mutations were lysed by
microfludization. The lysate was brought up to 30% ammonium
sulfate saturation, proteins were precipitated, and then resus-
pended in TED buffer. The remaining lysate was brought to 60%
ammonium sulfate saturation and proteins were precipitated and
resuspended, brought to 90% ammonium sulfate saturation and
precipitated and resuspended again. The proteins of interest were
present in the 60% and 90% ammonium sulfate precipitates.
These protein fractions were further purified by hydrophobic
interaction chromatography using a Phenyl FF 16/10 High Sub
column (GE Healthcare) followed by anion exchange chromatog-
raphy on a Source Q column (GE Healthcare) and size exclusion
chromatography on a Sephacryl S200 16/60 column (GE
Healthcare).

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pET24a_UDP_Xase_E162 was a gift from Sandra Gabelli (Addgene plasmid # 73160 ; http://n2t.net/addgene:73160 ; RRID:Addgene_73160)

  • For your References section:

    A UDP-X diphosphatase from Streptococcus pneumoniae hydrolyzes precursors of peptidoglycan biosynthesis. Duong-Ly KC, Woo HN, Dunn CA, Xu W, Babic A, Bessman MJ, Amzel LM, Gabelli SB. PLoS One. 2013 May 15;8(5):e64241. doi: 10.1371/journal.pone.0064241. Print 2013. PONE-D-13-06764 [pii] PubMed 23691178
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