| Title: | Alpha-factor leader sequence-directed transport of Escherichia coli beta-galactosidase in the secretory pathway of Saccharomyces cerevisiae |
| Author(s): | Das RC; Shultz JL; Lehman DJ; |
| Address: | "Research and Development, Miles Inc., Elkhart, IN 46515" |
| ISSN/ISBN: | 0026-8925 (Print) 0026-8925 (Linking) |
| Abstract: | "The construction of two fused genes is described. One involves the in-frame fusion of the yeast prepro-alpha-factor coding sequence, and the Escherichia coli lac Z gene. The second gene fusion utilizes a 103 bp yeast invertase NH2-terminal coding sequence at the fusion junction of the hybrid gene described above. The gene fusions, under the control of the alpha-factor promoter, expressed active beta-galactosidase in alpha haploid yeast cells. The activity could be regulated in a temperature-sensitive sir3 mutant. The incorporation of the invertase coding sequence at the MF alpha 1-lacZ fusion junction provided significantly higher levels of beta-galactosidase activity. A substantial quantity of the hybrid proteins generated from the gene fusions was primarily localized in the intracellular membranes of yeast cells, while a processed form could be secreted into the periplasm" |
| Keywords: | "Blotting, Northern Blotting, Western Cell Fractionation Cloning, Molecular Culture Media Escherichia coli/enzymology/*genetics Galactosidases/*genetics Gene Expression Regulation Mating Factor Peptides/*genetics Precipitin Tests Recombinant Fusion Protein;" |
| Notes: | "MedlineDas, R C Shultz, J L Lehman, D J eng Germany 1989/08/01 Mol Gen Genet. 1989 Aug; 218(2):240-8. doi: 10.1007/BF00331274" |
