Yeast harbor a single canonical G protein signaling system, the pheromone response pathway, responsible for the signal transduction of peptide mating pheromones that are secreted and exchanged between haploid yeast cells of opposite mating types. As with mammalian G protein signaling systems, the binding...
1995. pWITCH: a versatile two-hybrid assay vector for the production of epitope/activation domain-tagged proteins both in vitro and in yeast. Gene 165, 93–96. Article PubMed CAS Google Scholar Wong C., Naumovski L. 1997. Method to screen for relevant yeast two-hybrid-derived clones by...
The yeast two-hybrid system has provided a convenient means to both screen for proteins that interact with a protein of interest and to characterise the kn... DJ Stephens,G Banting - 《Traffic》 被引量: 93发表: 2000年 Using the yeast two‐hybrid assay to discover protein partners for the...
subtilis, lane 2: prepro signal peptide of mating factor α from S. cerevisiae, and lane 3: TFP1-4 in this study. hIL-2 was determined by an in vitro proliferation assay using the EL-4 (mouse T-lymphocyte) cell line (Fig. 3d). Recombinant yeast hIL-2 showed biological ...
To prepare the worms for the infection assay, the larvae were grown at 25°C for 4 days. This resulted in non-egg producing young adult worms. In order to maintain the worms, larvae were grown on at 15°C for 5 to 6 days until producing sufficient eggs for egg harvest. Method ...
Since aconitase is eclipsed distributed with minute amounts in the cytosolic fraction, we had to use the sensitive but qualitative α-complementation assay. The α-complementation assay employs the capability of two peptide fragments of β-galactosidase (designated ω-993 amino acids; α-77 amino ...
For a small percentage of baits, the repression assay does not work, although the bait protein is clearly present at high levels, and there is no reason to believe it is not nuclear. In these cases, it is generally reasonable to go ahead with the library screen. ...
We have initiated a systematic YTH screening program to identify kinase substrates in Arabidopsis and to investigate CDPK-mediated signal transduction pathways. In this study, we used constitutively active and/or catalytically impaired versions of AtCPK11 as baits in the YTH system to screen for At...
Peptides are loaded with a trap and elute configuration on C18 reverse-phase columns (ChIP C-18 precolumn 300 μm ID × 5 mm ChromXP and ChIP C-18 analytical column 75 μm ID × 15 cm ChromXP; Eksigent). - Peptides are eluted by using a 5–40% gradient of 0.1% (v/v) formic...
Both systems report disrupted interaction by a gain of signal which is easier to detect in a library screen as compared to a loss of signal. The Split-ubiquitin system (Figure 2D) was designed by Johnsson and Varshavsky in 1994 [53] to allow detection of protein-protein interactions occurring...