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重组肌红蛋白的原核表达、纯化及其冻干质控品的制备

, PP. 189-195

Keywords: 重组肌红蛋白,原核表达,纯化,冻干,质控品

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Abstract:

旨在为市场提供一种廉价、稳定、特异性强、灵敏度高的肌红蛋白质控品;将密码子优化后的肌红蛋白序列通过两步法进行化学合成,并连接到pET-28a表达载体上,转入大肠杆菌BL21(DE3),用0.5mmol/LIPTG37℃诱导表达;目的蛋白经NiSepharose6FastFlow(FF)亲和层析进行纯化后,对其特异性进行检测,并研究蛋白稀释液的配方和优化冻干工艺,最后对冻干质控品的稳定性进行检测;结果显示目的蛋白分子量大小为20kD左右,具有较强的特异性。蛋白冻干后具有很好的形态,37℃可以稳定保存10d;25℃、4℃可以稳定保存4个月以上。该蛋白的冻干制品成本低、稳定性好、特异性强、灵敏度高,可用于定性定量肌红蛋白检测试剂盒的质控品。

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