In vivo construction of cDNA libraries for use in the yeast two-hybrid system

We describe a simple and efficient one-step method to make cDNA libraries u sing homologous recombination in yeast. cDNA from any source, together with a linear vector, is used to transform yeast. Through homologous recombinat ion and gap repair, the cDNA is unidirectionally incorporated into the yeas t expression vector in vivo. The cDNA-encoded proteins can then be screened for potential protein-protein interactions with a bait already present in the yeast. This method allows rapid construction and screening of cDNA libr aries, even from extremely small amounts of mRNA, and can replace the use o f conventional cDNA libraries. Copyright (C) 1999 John Wiley & Sons, Ltd.