植物乳杆菌亚硝酸盐还原酶基因在大肠杆菌中表达  

Expression of nitrite reductase gene from Lactobacillus plantarum in Escherichia coli

作　　者：龚钢明[1] 何婷婷[1,2] 高能[3] 

机构地区：[1]上海应用技术学院,上海201418 [2]上海师范大学生命与环境科学学院,上海200234 [3]浙江理工大学生命科学学院,浙江杭州310018

出　　处：《中国酿造》2012年第11期135-137,共3页China Brewing

摘　　要：构建乳杆菌亚硝酸盐还原酶基因的重组质粒并表达。以植物乳杆菌基因组DNA为模板,PCR扩增亚硝酸盐还原酶基因,连接到表达载体pET-32a(+)上,构建的重组载体转入大肠杆菌BL21(DE3)中,经IPTG诱导表达后,菌体总蛋白经SDS-PAGE鉴定显示重组质粒诱导表达产物有特异性蛋白条带。该表达产物经细菌亚硝酸盐还原酶试剂盒测定显示有明显的酶活性。实验结果为深入研究植物乳杆菌亚硝酸盐还原酶提供有益的参考。The expression of nitrite reductase gene from Lactobacillus plantamm in Escherichia coli was studied. Nitrite reductase gene was amplified by PCR from the genomic DNA of L. plantarum, and then inserted into expression plasmid pET-32a （＋） to construct recombinant plasmid. The recombinant plasmid was transformed into E. coli BL21 （DE3）. The host bacteria containing recombinant plasmids was selected and grew with IPTG induction. The cell lysate of the host bacteria was extracted for SDS-PAGE and its enzymatic activity was determined. The results showed that the nitrite reductase gene was successfully expressed and the expressed protein contained nitrite reductase activity.

关 键 词：亚硝酸盐还原酶 植物乳杆菌 基因表达 

分 类 号：Q786[生物学—分子生物学]