Bacterial hypersecretion using mutant repressor sequence

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical

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435691, 4351723, 4352523, 43525233, 4353201, 435 91, 536 27, 935 11, 935 23, 935 29, 935 39, 935 41, 935 72, 935 73, C12P 2100, C12N 120, C12N 1500

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050305638

ABSTRACT:
An improved bacterial host cell useful for the inducible production and secretion in high yields of a heterologous protein is provided which contains a gene encoding the heterologous protein operatively linked to a secretory leader-encoding sequence and to an expression control sequence which contains a promoter region; and a second DNA sequence encoding a repressor capable of binding to said promoter region. The cell contains at least a mutation in the repressor binding region of the promoter or a mutation in the promoter binding region of the repressor-encoding sequence; or mutations in both regions. These mutation(s) lower the frequency of transcriptional induction by the promoter from the observed with the wild-type promoter and/or repressor-encoding sequence, resulting in higher yields of secreted heterologous protein.

REFERENCES:
Lambert et al., (1985), J. Bact., vol. 162, pp. 441-444.
Windle et al., (1986), vol. 95, pp. 95-99, Acne.
Hoffman et al., Pac Natl. Acad. Sci., vol. 82, p. 5107, 1985.
Ernst et al., Chem. Abst., vol. 107 (15), No. 128972a, "DNA Sequence, Multicopy Expression Vector, and Transformed Host for Enhanced Secretion of Heterologous Proteins Using Substituted Promoters".
Spicer et al., J. Biol. Chem., 050294321 257(10): 5716 (1982).

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