| 纳豆激酶基因在E.coli HB101中的初步表达研究 |
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| 引用本文： | 罗立新,黄志立,杨汝德,凌均建,梁世中. 纳豆激酶基因在E.coli HB101中的初步表达研究[J]. 微生物学通报, 2002, 29(3): 62-66 |
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| 作者姓名： | 罗立新 黄志立 杨汝德 凌均建 梁世中 |
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| 作者单位： | 1. 华南理工大学食品与生物工程学院,广州,510640
2. 深圳职业技术学院生物应用工程系 |
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| 基金项目： | 广东省自然科学基金资助项目 (No. 980 5 4 0 ) |
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| 摘 要： | 利用PCR技术以纳豆杆菌染色体DNA为模板扩增纳豆激酶基因，将该基因克隆到温度诱导型表达形体pVB220上，转化E．coliHB101，获得转豆激酶基因重组菌。在确定了其最佳培养时间与诱导时间后，SDS－PAGE分析结果表明基因表达产物为分泌型，蛋白表达量占菌体蛋白的12％左右，液体发酵后纳豆激酶产量可达120U／ml菌液，对重组菌中重组质粒的稳定性进行研究，结果表明该质粒在宿主菌中具有良好的分离稳定性，而结构稳定性较差。
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| 关 键 词： | 纳豆激酶 E.coliHB101 基因克隆 表达 质粒稳定性 |
| 文章编号： | 0253-2654(2002)03-0062-04 |
| 修稿时间： | 2001-04-25 |
CLONING OF NATTOKINASE GENE AND EXPRESSION IN E.COLI |
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| LUO Li-Xin HUANG Zhi-Li YANG Ru-De LING Jun-Jian LIANG Shi-Zhong. CLONING OF NATTOKINASE GENE AND EXPRESSION IN E.COLI[J]. Microbiology China, 2002, 29(3): 62-66 |
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| Authors: | LUO Li-Xin HUANG Zhi-Li YANG Ru-De LING Jun-Jian LIANG Shi-Zhong |
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| Abstract: | In this study, nattokinase gene was amplified by PCR using bacillus subtilis chromosomal DNA as template and cloned into expressed vector pBV220. After transforming recombinant plasmid into E.coli HB101, the recombinant strain was yielded. It was proved that expression products was secretive and expression protein was 12% of total cell protein by SDS-PAGE. Optimum culture time and inducing time was determined as 6h and 5h respectively. The plasmid stability studies showed that recombinant plasmid has excellent segregational stability but the structural stability was not good in the host cell. |
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| Keywords: | Nattokinase gene Cloning Expression Plasmid stability |
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