Combined transformation and genetic technique verification of protein-protein interactions in the yeast two-hybrid system

Shweta Tyagi, Sunil K. Lal

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16 Citations (Scopus)


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 raise 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. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)589-593
Number of pages5
JournalBiochemical and Biophysical Research Communications
Issue number3
Publication statusPublished - 2 Nov 2000
Externally publishedYes

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