Protocols and technical information

• Two-Hybrid vectors: pOBD2 and pOAD
• Recombination cloning
• Strains: PJ69-4a and PJ49-4alpha
• Yeast media

• PCR of genomic DNA (pdf)
• Expression of GST fusion proteins (pdf)
• GST pull-down (in vitro-binding assay, pdf)
• Lipid overlay assay (lipid-protein binding assay, pdf)

A selection of additional protocols can be found on the Herskowitz Lab Page.

Methods papers:

• Cagney,G. & Uetz,P. (2001) High-throughput two-hybrid screening using yeast arrays. Current Protocols in Protein Science 19.6: 1-13

• Cagney,G., Uetz,P. & Fields,S. (2000) High-throughput screening for protein-protein interactions using the two-hybrid assay. Meth. Enzymol. 328: 3-14

• Serebriskii, I.; Golemis, E. & Uetz, P. (2004) The yeast two-hybrid system for detecting interacting proteins. In: Walker, J.M. (ed.) The Proteomics Handbook, Humana Press, Totowa (NJ), (in press).

• Uetz,P., Ideker,T. & Schwikowski,B. (2002) Visualization and integration of protein-protein interactions [ 1.4 MB]. In: Golemis,E. (ed.) Protein-Protein Interactions - A Molecular Cloning Manual. Cold Spring Harbor Laboratory Press, pp. 623-646