eEF2-6xHis Expressing Yeast Strain TKY675

Yeast strain TKY675 expresses C-terminal 6xHis-tagged elongation factor 2 (eEF2) for rapid downstream purification.

Highlights

  • Yeast strain TKY675 is derived from the YPH499 background
  • Saccharomyces cerevisiae strain allows for expression of eEF2 possessing eukaryotic-specific protein modifications
  • C-terminal 6xHis-Tag allows for one-step downstream purification via nickel affinity chromatography

Elongation factors are proteins used in translation or protein synthesis during the process of cell cycle and elongation in some cells. In the ribosome, they facilitate translational elongation, from the formation of the first peptide bond to the formation of the last one. Specifically, elongation factor 2 catalyzes the GTP-dependent ribosomal translocation step during translation elongation. During this step, the ribosome changes from the pre-translocational (PRE) to the post-translocational (POST) state as the newly formed A-site-bound peptidyl-tRNA and P-site-bound deacylated tRNA move to the P and E sites, respectively. eEF2 catalyzes the coordinated movement of the two tRNA molecules, the mRNA and conformational changes in the ribosome.

From the laboratory of Terri Goss Kinzy, PhD, Rutgers University.

The Investigator's Annexe Part of The Investigator's Annexe program.

Catalog Number Product Size AVAILABILITY Price Qty
ED7004
eEF2-6xHis Expressing Yeast Strain TKY675
Spotted on filter disc In stock
Price: $321.00
Specifications
Product Type: Bacteria
Gene/insert name: Expresses S. cerevisiae eEF2 containing C-terminal 6xHis-Tag
Strain: Saccharomyces cerevisiae, TKY675 (Haploid)
Genotype: MATa ade2 leu2 ura3 his3 leu2 trp1 eft1::HIS3 eft2::TRP1 pEFT2-6xHis LEU2 CEN
Mating Type: MATa
Biosafety Level: 1
Growth Conditions: YEPD/30C
Format: Spotted on filter disc
Storage: Glycerol stock at -80C
Shipped: Ambient temperature
Documentation

Protocol Notes

Place disc on YEPD plate, add a few drops of sterile water to the disc and streak out.

Data

Expression and Purification of eEF2

Expression and Purification

(a) Yeast strains YEFD12h/pURA3-EFT1 (WT eEF2) and TKY675 (eEF2-6xHis) were grown overnight in YEPD to an A600 of 1.0, spotted as tenfold serial dilutions on YEPD medium and incubated for 2 d at either 303 (top panel) or 310 K (bottom panel). (b) Coomassie blue stained 10.8% SDS gel of eEF2-6xHis. Lane M, molecular-weight markers in kDa; lane 1, pool from Ni-chelating column.

Adapted from: Jørgensen, R, et al. Acta Crystallogr D Biol Crystallogr. 2002 Apr;58(Pt 4):712-5.

Provider
From the laboratory of Terri Goss Kinzy, PhD, Rutgers University.
References
  1. Jørgensen, R, Carr-Schmid, A, Ortiz, PA, Kinzy, TG, Andersen, GR. Purification and crystallization of the yeast elongation factor eEF2. Acta Crystallogr D Biol Crystallogr. 2002 Apr;58(Pt 4):712-5.

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