1887

Abstract

nuclease A hybrid genes, encoding proteins OmpA-nuclease, lipo-nuclease and Pin-nuclease, were cloned downstream of the yeast GAL 10 inducible promoter. OmpA-nuclease and lipo-nuclease contain the mature staphylococcal nuclease sequence preceded by the OmpA and lipoprotein signal sequences, respectively, whereas Pin-nuclease lacks a defined signal sequence at its amino terminus. We found that: () the nuclease gene products synthesized in yeast are active, but they do not affect cell growth; () OmpA-nuclease and lipo-nuclease are partially processed and constitute approximately 1·0–1·5% of the yeast cell protein; () OmpA and lipoprotein signal sequences function similarly in secretion, allowing 35–40% of the processed nuclease to be translocated into the yeast periplasm; and () Pin-nuclease, which lacks hydrophobic sequences at its amino-terminus, is accumulated at a level tenfold lower than the hybrid proteins that do contain signal sequences. Nevertheless, 50 % of the enzyme activity of Pin-nuclease in yeast is localized in the periplasmic space.

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1991-04-01
2021-04-16
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