The yeast shuttle/expression vector pORF-CLONE has been specifically developed for an improved and facilitated selection of cloned cDNA inserts containing open reading frames (ORFs) thus allowing an enriched growth of clones expressing authentic polypeptides. This vector system is particular useful for the development of a high-throughput technique for the one-step generation of high-quality cDNA libraries in the yeast Saccharomyces cerevisiae and a direct, time-saving screening of random-primed cDNA libraries. The selection system is based on the HIS3 marker gene fused to the C-terminus of the cDNA insert. The cDNAs cloned in-frame result in histidine prototrophic yeast cells growing on minimal medium, whereas clones bearing the vector without insert or out-of- frame inserts should not grow on this medium. |
