Cloning, expression and characterization of Kunming mice lactoferrin and its N-lobe

文献类型： 外文期刊

第一作者： Tian, Zigang

作者： Tian, Zigang;Teng, Da;Yang, Yalin;Wang, Jianhua;Wang, Jiarong;Tian, Zigang;Teng, Da;Yang, Yalin;Wang, Jianhua;Wang, Jiarong;Hu, Jiancheng

作者机构：

关键词： Escherichia coli;Expression;Kunming mice;Lactoferrin;N-lobe

期刊名称：BIOMETALS （ 影响因子：2.949； 五年影响因子：2.978 ）

ISSN：

年卷期：

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收录情况： SCI

摘要： The lactoferrin cDNA of Kunming mice was isolated by reverse transcription polymerase chain reaction and cloned into vector pET28a(+). Its deduced amino acid sequence was analyzed and compared with lactoferrin of other species. Its secondary and tertiary structure are predicted and modeled by bioinformatics tools online. Then recombinant Kunming mice lactoferrin and its N-lobe were both expressed successfully in the Escherichia coli BL21(DE3) in the form of inclusion bodies. After purification with Ni-NTA His-Bind resin, the yield of recombinant lactoferrin was 17 mg l~(-1) with purity of 92.1%, and that of lactoferrin N-lobe was 20 mg l~(-1) with purity of 98.5%. The inhibition efficiency of refolded lactoferrin N-lobe against Staphylococcus aureus ATCC 25923 reaches 48.6% at the concentration of 25 μmol l ~(-1). However, the refolded lactoferrin (12.5 μmol l~(-1)) didn't display obvious inhibition activity in the test. The expression of recombinant Kunming mice lactoferrin and its N-lobe will be helpful for the study of lactoferrin on structure, function and application in a mouse model system.

分类号： Q

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