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SMAP-29抗菌肽的原核表达、纯化及活性检测

, PP. 144-148

Keywords: 抗菌肽,SMAP-29,原核表达,抗菌活性

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Abstract:

旨在利用基因工程技术表达出有活性的重组SMAP-29抗菌肽。根据大肠杆菌偏好的密码子优化smap-29基因序列,在目标肽序列N端添加肠激酶识别位点,C端添加终止密码子,化学合成基因序列,通过EcoRI和HindIII双酶切位点连接到pET-28a(+)上构建重组表达载体,在大肠杆菌BL21(DE3)中表达重组蛋白,Ni-NTA亲和层析纯化,肠激酶切割后释放出SMAP-29抗菌肽,检测其抗菌活性。结果显示,带有6×His标签及肠激酶识别位点的SMAP-29重组融合蛋白在大肠杆菌中以包涵体形式表达,利用Ni-NTA亲和层析可获得纯化的融合蛋白,肠激酶切割后释放出的SMAP-29重组抗菌肽对大肠杆菌、金黄色葡萄球菌和白念珠菌的最小抑菌浓度依次为10、20和40μmol/L。

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