Cloning, characterization and application of the promoter region of the alkaline protease gene in <I>Bacillus alcalophillus</I> PB92

Abstract

Promoter function fragment of alkaline protease gene (GenBank accession number: EU130686) was cloned from Bacillus alcalophillus PB92 genome by TAIL-PCR. Sequenced and analyzed revealed that it contains several typical promoter characterized regions. Two reverse translation frames were located in -538~-370 bp and-275~-128 bp. Deletion analysis of the sequence demonstrated that 414 bp to 619 bp upstream of the TSS showed predominant promoter activity, and a 105 bp length sequence can serve as this function. Additionally, a representative Sec-type signal peptide structure was detected in PB92 AprE signal peptide. By cloning the PaprE and alkaline protease signal peptide gene into pBE2, an expression vector pBEAC was constructed, and a plant sweet protein monellin gene was highly expressed in B. subtilis 1A751.