| 一种新型IIa类细菌素的克隆表达和活性鉴定 |
| |
| 引用本文： | 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. |
| |
| 作者姓名： | 谢燕 陈海琴 张秋香 田丰伟 陈永泉 张灏 陈卫 |
| |
| 作者单位： | 江南大学食品学院，无锡，214122 |
| |
| 基金项目： | 国家自然科学基金 (Nos. 20706023, 20836003) 资助。 |
| |
| 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。
|
| 关 键 词： | IIa类细菌素，融合表达，蛋白质纯化，活性鉴定 |
| 收稿时间： | 9/6/2010 12:00:00 AM |
Expression and characterization of a new class IIa bacteriocin |
| |
| Yan Xie,Haiqin Chen,Qiuxiang Zhang,Fengwei Tian,Yongquan Chen,Hao Zhang and Wei Chen.Expression and characterization of a new class IIa bacteriocin[J].Chinese Journal of Biotechnology,2011,27(7):976-982. |
| |
| Authors: | Yan Xie Haiqin Chen Qiuxiang Zhang Fengwei Tian Yongquan Chen Hao Zhang and Wei Chen |
| |
| Institution: | School of Food Science and Technology, Jiangnan University, Wuxi 214122, China;School of Food Science and Technology, Jiangnan University, Wuxi 214122, China;School of Food Science and Technology, Jiangnan University, Wuxi 214122, China;School of Food Science and Technology, Jiangnan University, Wuxi 214122, China;School of Food Science and Technology, Jiangnan University, Wuxi 214122, China;School of Food Science and Technology, Jiangnan University, Wuxi 214122, China;School of Food Science and Technology, Jiangnan University, Wuxi 214122, China |
| |
| Abstract: | 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. |
| |
| Keywords: | class IIa bacteriocin fusion expression protein purification activity identification |
| 本文献已被 CNKI 万方数据 PubMed 等数据库收录！ |
| 点击此处可从《生物工程学报》浏览原始摘要信息 |
| 点击此处可从《生物工程学报》下载免费的PDF全文 |
|
