The structural basis of the high in vivo strength of the rRNA P2 promoter of Escherichia coli.
Ribosomal RNA promoters of Escherichia coli are probably the strongest promoters in vivo and they can be used on plasmid vectors to express protein-coding sequences at a high rate. In fact, the P2 promoter of the rrnB gene is stronger (in vivo) than the tac promoter, which has a perfect consensus sequence. Conversion of the rrnB P2 promoter sequence to consensus significantly increases in vivo promoter strength. The removal of four nucleotides downstream of the -10 region also increases the strength of this promoter. On the other hand, shifting of the A + T-rich region upstream of this promoter by an 11-bp insertion drastically decreases in vivo activity. It is concluded that the two functionally important hexanucleotide sequences, -35 and -10, are necessary but not sufficient factors for the optimalization of in vivo promoter strength.
Lukacsovich
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Institute of Biochemistry, Biological Research Center, Szeged, Hungary.
Gaal
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Venetianer
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D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics*; D007763:Lac Operon; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D012335:RNA, Ribosomal / Q000235:genetics*; D015183:Restriction Mapping; D014158:Transcription, Genetic
D012335:RNA, Ribosomal