GST-颗粒溶解素融合蛋白的纯化及其抗菌活性分析

闵宏林; 沈继龙; 陈勇; 李柏青

GST-颗粒溶解素融合蛋白的纯化及其抗菌活性分析

1. 安徽医科大学病原生物学教研室, 安徽合肥 230032;
2. 蚌埠医学院病原生物学教研室, 安徽蚌埠 233030;
3. 蚌埠医学院免疫学教研室, 安徽省感染与免疫重点实验室(蚌埠医学院), 安徽蚌埠 233030

基金项目:

安徽省自然科学基金资助项目(050430603)

安徽省教育厅自然科学研究资助项目(2006kj122zc)

详细信息

作者简介:
闵宏林(1971-),女,讲师.

中图分类号: R378.21;R392.11
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- 收稿日期: 2006-09-19

Purification and antibacterial activity analysis of GST-granulysin fusion protein

1. Department of Microbiology, Anhui Medical University, Hefei 230032;
2. Department of Microbiology, Anhui Key Laboratory of Infection and Immunity, Bengbu Medical College, Bengbu 230030, China;
3. Department of Immunology, Anhui Key Laboratory of Infection and Immunity, Bengbu Medical College, Bengbu 230030, China

摘要

摘要: 目的: 纯化大肠埃希菌表达的GST-颗粒溶解素融合蛋白,并分析其体外抗菌活性。方法: 将IPTG诱导处理后的细菌破碎,分离含有可溶性GST-颗粒溶解素融合蛋白的细菌裂解液,并通过GSH-琼脂糖珠亲和层析柱纯化。同时采用MTT法分析该纯化蛋白的抗菌活性。结果: GST-颗粒溶解素融合蛋白能以可溶性形式在大肠埃希菌中表达,采用亲和层析法可获得分子量为44kDa的单一蛋白条带。经抗菌活性分析,表明纯化产物对金黄色葡萄球菌和甲型溶血性链球菌具有显著抑制作用,而对伤寒沙门菌和大肠埃希菌BL21的抗菌活性不明显。结论: 成功纯化GST-颗粒溶解素融合蛋白,该蛋白对革兰阳性菌具有高效抗菌活性。
- 大肠埃希菌 /
- 颗粒溶解素 /
- 融合表达 /
- 纯化 /
- 抗菌作用
Abstract: Objective: To purify GST-granulysin fusion protein expressed in E.coli and identify its antibacterial activity in vitro.Methods: E.coli was sonicated after it was induced by IPTG,and GST-granulysin fusion protein was separated from the soluble extract.The fusion protein was purified by affinity chromatography with glutathione agarose.Its bactericidal activity was analysed by MTT assay.Results: Expression of GST-granulysin in E.coli was detected by SDS-page methods.After affinity chromatography,a single protein band at about molecular weight 44 kDa was obtained.The result of bactericidal activity analysis indicated that it showed potent antibacterial activity against Staphylococcus aureus and Streptococcus sanguis,but it showed weaker antibacterial activity against Salmonella enterica and E.coli BL21.Conclusions: The GST-granulysin fusion protein was successfully purified,and it exhibits effective bactericidal activity against gram positive bacteria.
- Escherichia coli /
- granulysin /
- fusion expression /
- purification /
- bactericide

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文章访问数: 2847
HTML全文浏览量: 265
PDF下载量: 16
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GST-颗粒溶解素融合蛋白的纯化及其抗菌活性分析

作者简介: 闵宏林(1971-),女,讲师.

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Purification and antibacterial activity analysis of GST-granulysin fusion protein

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作者简介:
闵宏林(1971-),女,讲师.