News & Highlights: Hutchinson-Gilford progeria syndrome (HGPS) is a rare and fatal disease characterized by premature aging. In their recent collaboration, YRC researcher John Yates and collaborator Juan Carlos Izpisua Belmonte found induced pluripotent stem cells from HGPS patients lacked molecular characteristics associated with the disease, which were restored upon differentiation. See their paper in Nature to learn more. [Read Article]
News & Highlights: YRC researcher Stan Fields used the model organism Saccharomyces cerevisiae to probe the effects of nutritionally acquired metabolites on statins, a cholesterol-lowering drug widely prescribed to prevent heart disease. He found that copper and zinc ions impair the effect of statins by upregulating genes related to sterol production. Please read his paper in Molecular BioSystems to learn more. [Read Article]
News & Highlights: YRC researchers David Baker and Stan Fields have developed new technology for examining how a protein's sequence affects its function. This new technology is large-scale and may be applied to many in vitro or in vivo protein assays, providing a general means for studying the functional consequences of protein variation. Please read their paper in Nature Methods to learn more. [Read Article]
News & Highlights: The YRC collaborated with Sue Biggins at the Fred Hutchinson Cancer Research Center in Seattle to examine centromeres, whose proper function is critical to prevent conditions associated with cancer and some birth defects. This work, performed in yeast, was recently published in Molecular Cell, where Dr. Biggins proposes a new pathway for the regulation of centromeric function. [Read Article]
News & Highlights: Multidimensional protein identification technology (MudPIT) developed by the YRC was used in a recent collaboration with David Drubin at the University of California, Berkeley, to examine the assembly of actin networks in yeast. In his recent paper in Current Biology, Dr. Drubin describes the nucleation and assembly of these large protein complexes, and how MudPIT was used to characterize their composition. [Read Article]

Cloning vectors (pOBD2)

DNA binding domain vector: pOBD2 (with uniques restriction sites and main features

Sequence features:

  • DH1 promoter: 1-298 (at least; region of homology when searched against SGD)
  • GAL4-DBD: 434-982
  • ADH1 terminator: 1221-1687 (at least; region of homology to SGD genom. seq.)
  • beta lactamase (amp resistance): 4363-5220
  • TRP1: 3379-4050
  • CEN4: 1814-2667

Cloning site (position 877-1012):

C TAT CTA TTC GAT GAT GAA GAT ACC CCA CCA AAC CCA AAA AAA GAG ATC GAA TTC CAG CTG ACC ACC ATG GCA ATT CCC GGG GAT CCG TCG ACC TGC AGA GAT CTA TGA ATC GTA GAT ACT GAA AAA CCC CGC AAG
This sequence corresponds exactly to the two re-PCR primers (green + blue = forward primer, pink + black = reverse primer). The two restriction sites for linearizing pOBD2 (PvuII = CAGCTG and NcoI = CCATGG) are underlined. In the 4 bp-region of mixed color (blue and pink) the two primer sequences overlap).
The green and blue sequences here correspond to the red and grey sequences in the figure below. Pink and black correspond to beige and grey (sorry for the confusing color codes).

Actual sequence of pOBD2 Plasmids & Protocols