Combined transformation and genetic technique verification of protein-protein interactions in the yeast two-hybrid system.
The yeast two-hybrid system is frequently used to identify protein-protein interactions. The assay is based on the functional reconstitution of a transcriptional activator. Since an indirect phenotype of the positive clones is the basis for selection of positive interacting clones, the two-hybrid screens are vulnerable to false positives. Here we report a screening protocol based on the sequential use of the cotransformation approach followed by the genetic method for verifying true two-hybrid interactions. Using this procedure, we have screened a cDNA library and have been able to isolate true positives from the yeast two-hybrid screen.
Tyagi
S
S
Virology Group, International Centre for Genetic Engineering and Biotechnology, New Delhi, 1100069, India.
Lal
S K
SK
D015723:Gene Library; D010641:Phenotype; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014170:Transformation, Genetic / Q000235:genetics*; D020798:Two-Hybrid System Techniques