Nucleotide sequence and cloning in Bacillus subtilis of the Bacillus stearothermophilus pleiotropic regulatory gene degT.

摘要: The regulatory gene (degT) from Bacillus stearothermophilus NCA1503 which enhanced production of extracellular alkaline protease (Apr) was cloned in subtilis with pTB53 as a vector. When B. MT-2 (Npr- [deficiency neutral protease] Apr+) transformed the recombinant plasmid, pDT145, plasmid carrier produced about three times more than did wild-type strain. In contrast, when DB104 Apr-) used host, transformant pDT145 could not exhibit any activity. After construction deletion plasmids, DNA sequencing done. A large open reading frame found, and nucleotide sequence analysis showed that degT composed 1,116 bases (372 amino acid residues, molecular weight 41,244). Shine-Dalgarno found nine upstream frame. strain carrying following pleiotropic phenomena: (i) enhancement enzymes such levansucrase, (ii) repression autolysin activity, (iii) decrease transformation efficiency for (competent cell procedure), (iv) altered control sporulation, (v) loss flagella, (vi) abnormal division. SIC1 degT, transformants exhibited These phenomena are similar to those phenotypes degSU(Hy) (hyperproduction), degQ(Hy), degR mutants subtilis. However, product (DegT) is different reported products. Furthermore, DegT includes hydrophobic core region N-terminal portion (amino numbers 50 160), consensus binding 160 179), homologous transcription activator proteins 351 366). We discuss possibility membrane protein functions sensor transfers signal environmental stimuli target genes activate or repress genes. Images