Alpha-factor leader sequence-directed transport of Escherichia coli beta-galactosidase in the secretory pathway of Saccharomyces cerevisiae |
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Authors: | Rathin C Das Janice L Shultz and Donna J Lehman |
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Institution: | (1) Research and Development, Miles Inc., 46515 Elkhart, IN, USA;(2) Present address: Bayer AG, PH-P VE Biochemie, D-5600 Wuppertal 1, Federal Republic of Germany |
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Abstract: | Summary The constuction of two fused genes is described. One involves the in-frame fusion of the yeast prepro--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 -factor promoter, expressed active -galactosidase in haploid yeast cells. The activity could be regulated in a temperature-sensitive sir3 mutant. The incorporation of the invertase coding sequence at the MF1-lacZ fusion junction provided significantly higher levels of -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.A portion of this work appeared in Biotechnology Progress (Das and Shultz 1986) as proceedings of the symposium on Industrial Scale Protein Purification, held at the annual meeting of the Institute of Chemical Engineers in Miami Beach, Fla, USA on November 4, 1986 |
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Keywords: | Yeast MF1 leader" target="_blank">gif" alt="agr" align="BASELINE" BORDER="0">1 leader Gene fusion Secretion |
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