NOVEL PROMOTER PROBE VECTOR PRODUCED BY GENE RECOMBINATION AND NOVEL PROMOTER PRODUCED BY USING SAID VECTOR

摘要

PURPOSE:To enable cloning of a promoter which has hitherto been difficult, by integrating a specific gamma-glutamyl-L-cystine synthetic enzyme gene in a vector replicable in a yeast belonging to Saccharomyces genus, etc. CONSTITUTION:Plasmid pGSR-100 is cut with restriction enzyme ScaI and partially cut with HpaI to obtain a DNA fragment containing complete GSH-I gene and BamHI linker is attached to both ends of the fragment. beta-Galactosidase gene is removed from pMC-1585 and the above DNA fragment is inserted to the BamHI incision site of the obtained plasmid pGSR-I. The BamHI incision site at the downstream side of GSRH-I gene on the pGSR-150 is removed by using T4 polymerase to obtain a plasmid 151. A DNA fragment (P-S) expressing promoter activity in yeast is inserted to the BamHI site upstream of the plasmid 151.