枯草芽孢杆菌apr基因的克隆和表达

通过PCR技术克隆枯草芽孢杆菌蛋白酶apr基因,并分别对该基因和编码蛋白进行同源性比较和酶学性质分析。结果表明,apr基因序列全长1509bp,编码503个氨基酸,同源性100%;克隆到表达质粒pET-22b(+)后,将重组质粒经热激转化至大肠杆菌BL21(DE3)中,IPTG诱导表达,蛋白分子质量为55ku。测定酶活为19500U·mL-1。该基因克隆和表达的成功对于制备大豆抗氧化肽具有实际应用价值,对进一步深入研究其生物学和酶学机制具有重要意义。

Clone and expression of Bacillus subtilis apr gene

Bacillus subtilis protease apr gene was cloned by PCR technology,the homology of this gene and code protein were compared,which enzymology characteristic was analyzed.It showed that full-length of gene sequence was 1 509 bp,503 amino acid being coded,and the consistency of sequencing results was 100%.Recombinant expression plasmid pET-22b(+)-apr was constructed and expressed in E.coli BL21 successfully,protein molecular weight was 55 ku,and the determined enzyme activity was 19 500 U·mL-1.The cloning and analysis of this gene could provide valuable gene material for small peptide preparing of soybean protein,which had great significance on deep studying of the molecular biologicalmechanism.