Yeast Resource Center | Strains and Plasmids
15868
page-template-default,page,page-id-15868,ajax_fade,page_not_loaded,,qode-theme-ver-10.1.2,wpb-js-composer js-comp-ver-5.1,vc_responsive

Strains and Plasmids

How To Request Strains or Plasmids

A number of plasmids and strains are available through the YRC for two-hybrid analysis (see below). For plasmid requests you must select whether they will be used for commercial or non-commercial purposes. Also, specify how you would like us to send the plasmids (FedEx or standard post), and provide a FedEx account number for billing if you choose FedEx.

All requests for plasmids or strains must be submitted through our collaborators’ website. Use the log in form to the right to log into the site, or the registration button if you are a new collaborator.

Yeast Two-Hybrid Strains

Gal4-DNA-binding domain fusions (“baits”) are expressed in PJ69-4alpha:

MATalpha trp1-901 leu2-3,112 ura3-52 his3-200 gal4(deleted) gal80(deleted) LYS2::GAL1-HIS3 GAL2-ADE2 met2::GAL7-lacZ

GAL4 activation domain fusions (“preys”) are expressed in PJ69-4a:

MATa trp1-901 leu2-3,112 ura3-52 his3-200 gal4(deleted) gal80(deleted) LYS2::GAL1-HIS3 GAL2-ADE2 met2::GAL7-lacZ

Both strains are identical except for the mating type. PJ69-4a was constructed by Phil James et al. (1996) and is described in:

James, P., Halladay, J. & Craig, E. A. (1996) Genomic libraries and a host strain designed for highly efficient two- hybrid selection in yeast. Genetics 144: 1425-1436

Yeast Two-Hybrid Plasmids

Please see the Fields Lab website for descriptions of cloning vectors pOBD2 and pOAD. Please also see the Fields Lab website for descriptions of PCR primers for amplification of ORFs and re-PCRs.

Fluorescence Microscopy Plasmids

We have created a number of different plasmids that are useful for tagging proteins with CFP, YFP, or DsRed. These plasmids can be used for fusions to either the N or C terminus of your protein of interest. We also supply a plasmid to create a positive control for FRET. These plasmids are available through addgene, a nonprofit plasmid repository.

 

C-terminal tags (Wach)
  • Venus, G418 selection (pBS7) (Nagai)
  • YFP, his selection (pDH5)
  • YFP, G418 selection (pDH6)
  • CFP, G418 selection (pDH3)
  • CFP, hygromycin-B selection (pBS4) (Goldstein)
  • DsRed.T1.N1, G418 selection (pTY24)
  • Cerulean, hygromycin-B selection (pBS10)
  • mCherry, G418 selection (pBS34)
  • mCherry, hygromycin-B selection (pBS35)
N-terminal tags (Prein)
  • YFP, G418 selection (pDH22)
  • CFP, G418 selection (pBS5)
  • Venus, G418 selection (pBES2)
  • Cre recombinase (pSH47)
FRET positive control
  • YFP-CFP (his selection) (pDH18)
  • YFP-CFP (G418) [pFA6a-YFP-CFP::KanR]