Xie Yan, Chen Haiqin, Zhang Qiuxiang, Tian Fengwei, Chen Yongquan, Zhang Hao, Chen Wei
School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.
Sheng Wu Gong Cheng Xue Bao. 2011 Jul;27(7):976-82.
NB-Cl gene is a potential class IIa bacteriocin gene. To obtain its soluble expression, NB-C1 was fused with the green fluorescent protein (GFP) gene and a recombinant expression vector plVEX 2.4d-GFP-NB-C1 was constructed, which was transformed into Escherichia coli BL21(DE3) pLysS. The expressed fusion protein GFP-NB-CI was purified by Ni-NTA affinity chromatography and the bioactivity was examined using Listeria monocytogenes as the indicator bacteria. The results showed that the expressed fusion protein GFP-NB-C1 was soluble and the final concentration of the purified fusion protein was 36.1 mg/L E. coli culture and had the purity above 95%. The antimicrobial assay of GFP-NB-C1 was analyzed and showed its high activity against Listeria monocytogenes.
NB-Cl基因是一种潜在的IIa类细菌素基因。为实现其可溶性表达,将NB-C1与绿色荧光蛋白(GFP)基因融合,构建了重组表达载体pLVEX 2.4d-GFP-NB-C1,并将其转化至大肠杆菌BL21(DE3)pLysS中。通过Ni-NTA亲和层析法纯化表达的融合蛋白GFP-NB-CI,并以单核细胞增生李斯特菌作为指示菌检测其生物活性。结果表明,表达的融合蛋白GFP-NB-C1是可溶的,纯化后的融合蛋白在大肠杆菌培养物中的终浓度为36.1 mg/L,纯度高于95%。对GFP-NB-C1的抗菌活性进行分析,结果显示其对单核细胞增生李斯特菌具有高活性。