MINIMUM LENGTH OF SEQUENCE HOMOLOGY REQUIRED FOR IN-VIVO CLONING BY HOMOLOGOUS RECOMBINATION IN YEAST

With efficient homologous recombination in Saccharomyces cerevisiae, a rapid in vivo cloning technique has been available. Here we demonstra ted that 30 bp of a homologous sequence at each end of a DNA fragment is sufficient to integrate the fragment into a linearized plasmid in y east. To obtain a high yield of recombination transformants, however, more than 60 bp are desirable. Interestingly, we observed that 20 bp o f homology at one end of a DNA fragment is sufficient far efficient re combination provided that the other end contains SO bp of homologous s equence, Some applications, including high-throughput transferring of EST inserts to the yeast expression systems for the Human Genome Proje ct, are discussed. (C) 1997 Academic Press.