1887

Abstract

The two promoters and of , and the promoter of were investigated with respect to growth-phase-dependent expression and regulation in transformants using the reading frame encoding BgaH, an enzyme with -galactosidase activity, as reporter. For comparison, the promoter of the ferredoxin gene of and the promoter of (formerly ) were analysed. , driving the expression of a house-keeping gene, was highly active during the exponential growth phase, whereas and the three promoters yielded the largest activities during the stationary growth phase. Compared to , the basal promoter activities of and were rather low, and larger activities were only detected in the presence of the endogenous transcriptional activator protein GvpE. The promoter does not yield a detectable basal promoter activity and is only active in the presence of the homologous cGvpE. To investigate whether the -TATA box and the BRE element were the reason for the lack of the basal activity, these elements and also sequences further upstream were substituted with the respective sequences of the stronger promoter and investigated in transformants. All these promoter chimera did not yield a detectable basal promoter activity. However, whenever the -BRE element was substituted for the -BRE, an enhanced cGvpE-mediated activation was observed. The promoter chimeras harbouring -BRE plus 5 (or more) bp further upstream also gained activation by the heterologous pGvpE and mcGvpE proteins. The sequence required for the GvpE-mediated activation was determined by a 4 bp scanning mutagenesis with the 45 bp region upstream of -BRE. None of these alterations influenced the basal promoter activity, but the sequence TGAAACGG-n4-TGAACCAA was important for the GvpE-mediated activation of .

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2005-01-01
2019-10-17
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