一种新型IIa类细菌素的克隆表达和活性鉴定
Expression and characterization of a new class IIa bacteriocin
投稿时间:2010-09-06  
DOI:  
中文关键词:IIa类细菌素,融合表达,蛋白质纯化,活性鉴定
英文关键词:class IIa bacteriocin, fusion expression, protein purification, activity identification
基金项目:国家自然科学基金 (Nos. 20706023, 20836003) 资助。
作者单位E-mail
谢燕 江南大学食品学院无锡 214122  
陈海琴 江南大学食品学院无锡 214122  
张秋香 江南大学食品学院无锡 214122  
田丰伟 江南大学食品学院无锡 214122  
陈永泉 江南大学食品学院无锡 214122  
张灏 江南大学食品学院无锡 214122  
陈卫 江南大学食品学院无锡 214122 weichen@jiangnan.edu.cn 
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中文摘要:
      NB-C1为一种潜在的IIa类细菌素基因,为实现其在大肠杆菌中的高效可溶表达,首先构建了NB-C1蛋白与绿色荧光蛋白 (GFP) 的融合表达载体pIVEX 2.4d-GFP-NB-C1,然后将构建的表达载体转化大肠杆菌BL21(DE3) pLysS,经诱导表达后,重组蛋白GFP-NB-C1以可溶的形式存在于细胞内。经Ni-NTA亲和层析柱分离纯化后,重组融合蛋白的纯度大于95%,产量达36.1 mg/L。抑菌试验表明,纯化后的重组蛋白对单核细胞增生李斯特氏菌具有明显的抑制作用。
英文摘要:
      NB-C1 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 pIVEX 2.4d-GFP-NB-C1 was constructed, which was transformed into Escherichia coli BL21(DE3) pLysS. The expressed fusion protein GFP-NB-C1 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.
谢燕,陈海琴,张秋香,田丰伟,陈永泉,张灏,陈卫.一种新型IIa类细菌素的克隆表达和活性鉴定[J].生物工程学报,2011,27(7):976~982
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