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纳豆激酶基因克隆及其在大肠杆菌中活性表达研究

闫达中,许芳,李洁,冯建成,杨艳燕

JOURNAL OF HUBEI UNIVERSITY（NATURAL SCIENCE EDITION）（2003）

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Abstract
以纳豆芽孢杆菌基因组DNA为模板,PCR扩增了纳豆激酶基因(natto kinase gene)中编码前肽、成熟肽的核苷酸序列(pro-NK),构建大肠杆菌表达质粒pTYB102,转化大肠杆菌ER2566.在IPTG诱导下,分别在 15℃(14?h)、30℃(3h)、37℃(2h)条件下培养,pTYB102均能表达出有活性的纳豆激酶.实验证实纳豆激酶基因得到活性表达需要Pro序列.SDS-PAGE表明,15℃比30℃和37℃培养表达的杂蛋白更少.薄层扫描测定表达的纳豆激酶占菌体总蛋白的30%以上.
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