Abstract
The signal sequences of the MFα1 prepro α-factor gene are frequently used to direct secretion of heterologous proteins fromSaccharomyces cerevisiae. They are often employed together with the MFα1 promoter in secretion vectors, such that this promoter directs the transcription of many heterologous gene cassettes in yeast. Most of the existing literature indicates that the MFα1 promoter is constitutive in MATα cells, although some data suggests that it may be more active in respiratory or late logarithmic fermentative cultures. To identify whether there is a growth rate or medium control over MFα1 promoter activity a strain was constructed with an integrated MFα1 promoter β-galactosidase (lacZ) reporter gene fusion. Intracellular β-galactosidase of this strain during batch culture on glucose, raffinose and acetate showed that MFα1 promoter activity was higher during respiratory growth on acetate as compared to more rapid fermentative growth on glucose or raffinose a result that might indicate this activity being inversely related to growth rate. Chemostat culture confirmed that growth rate does indeed influence MFα1 promoter activity in glucose-grown cells, the activity of this promoter increasing 2- to 2.5-fold as dilution (growth) rates were reduced from maximal values to 0.2 h−1, but then decreasing with the further decreases in dilution rate needed for fully respiratory growth. Thus a promoter generally thought to be constitutive in MATα cells is nevertheless subject to a complex growth rate control.
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Kirk, N., Piper, P.W. Growth rate influences MFα1 promoter activity in MATαSaccharomyces cerevisiae . Appl Microbiol Biotechnol 42, 340–345 (1994). https://doi.org/10.1007/BF00902739
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DOI: https://doi.org/10.1007/BF00902739