Expression of nattokinase in E. coli. and preparation of its polyclonal antibody(纳豆激酶原核表达纯化及多克隆抗体的制备)
纳豆激酶是一种源于日本传统食品纳豆且具有强烈溶栓作用的丝氨酸蛋白酶.本研究将编码信号肽、前导肽和成熟肽在内的纳豆激酶基因克隆到原核表达载体pET28a+中，获得重组表达质粒pETNK.将此重组表达栽体质粒转化到大肠杆菌BL21中，经IPTG诱导表达.SDS-PAGE结果显示纳豆激酶在大肠杆菌中成功表达，表达的纳豆激酶融合蛋白分子量大约为31kD.亲和层析法纯化表达产物，并以此纯化产物作为免疫原免疫新西兰大白兔，制备兔抗纳豆激酶血清，用ELISA和Western blotting对制备的兔抗血清中的多克隆抗体进行了鉴定，纤维蛋白平板实验显示，表达的纳豆激酶融合蛋白具有较好的溶纤活性....
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| Format: | Article |
| Language: | zho |
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Zhejiang University Press
2006-07-01
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| Series: | Zhejiang Daxue xuebao. Lixue ban |
| Subjects: | |
| Online Access: | https://doi.org/zjup/1008-9497.2006.33.4.447-450 |
| _version_ | 1827303401854074880 |
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| author | ZHULi-cheng(朱立成) ZHANGYao-zhou(张耀洲) |
| author_facet | ZHULi-cheng(朱立成) ZHANGYao-zhou(张耀洲) |
| author_sort | ZHULi-cheng(朱立成) |
| collection | DOAJ |
| description | 纳豆激酶是一种源于日本传统食品纳豆且具有强烈溶栓作用的丝氨酸蛋白酶.本研究将编码信号肽、前导肽和成熟肽在内的纳豆激酶基因克隆到原核表达载体pET28a+中，获得重组表达质粒pETNK.将此重组表达栽体质粒转化到大肠杆菌BL21中，经IPTG诱导表达.SDS-PAGE结果显示纳豆激酶在大肠杆菌中成功表达，表达的纳豆激酶融合蛋白分子量大约为31kD.亲和层析法纯化表达产物，并以此纯化产物作为免疫原免疫新西兰大白兔，制备兔抗纳豆激酶血清，用ELISA和Western blotting对制备的兔抗血清中的多克隆抗体进行了鉴定，纤维蛋白平板实验显示，表达的纳豆激酶融合蛋白具有较好的溶纤活性. |
| first_indexed | 2024-04-24T17:01:31Z |
| format | Article |
| id | doaj.art-5b5c4c88a13749a3a3a1dd246eda203e |
| institution | Directory Open Access Journal |
| issn | 1008-9497 |
| language | zho |
| last_indexed | 2024-04-24T17:01:31Z |
| publishDate | 2006-07-01 |
| publisher | Zhejiang University Press |
| record_format | Article |
| series | Zhejiang Daxue xuebao. Lixue ban |
| spelling | doaj.art-5b5c4c88a13749a3a3a1dd246eda203e2024-03-29T01:58:23ZzhoZhejiang University PressZhejiang Daxue xuebao. Lixue ban1008-94972006-07-01334447450zjup/1008-9497.2006.33.4.447-450Expression of nattokinase in E. coli. and preparation of its polyclonal antibody(纳豆激酶原核表达纯化及多克隆抗体的制备)ZHULi-cheng(朱立成)0ZHANGYao-zhou(张耀洲)1 1.College of Life Sciences, Zhejiang University, Hangzhou 310029, China( 1.浙江大学生命科学学院，浙江 杭州 310029) 2.College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China( 2.浙江理工大学生命科学学院，浙江 杭州 310018)纳豆激酶是一种源于日本传统食品纳豆且具有强烈溶栓作用的丝氨酸蛋白酶.本研究将编码信号肽、前导肽和成熟肽在内的纳豆激酶基因克隆到原核表达载体pET28a+中，获得重组表达质粒pETNK.将此重组表达栽体质粒转化到大肠杆菌BL21中，经IPTG诱导表达.SDS-PAGE结果显示纳豆激酶在大肠杆菌中成功表达，表达的纳豆激酶融合蛋白分子量大约为31kD.亲和层析法纯化表达产物，并以此纯化产物作为免疫原免疫新西兰大白兔，制备兔抗纳豆激酶血清，用ELISA和Western blotting对制备的兔抗血清中的多克隆抗体进行了鉴定，纤维蛋白平板实验显示，表达的纳豆激酶融合蛋白具有较好的溶纤活性.https://doi.org/zjup/1008-9497.2006.33.4.447-450纳豆激酶原核表达抗体制备纤溶活性 |
| spellingShingle | ZHULi-cheng(朱立成) ZHANGYao-zhou(张耀洲) Expression of nattokinase in E. coli. and preparation of its polyclonal antibody(纳豆激酶原核表达纯化及多克隆抗体的制备) Zhejiang Daxue xuebao. Lixue ban 纳豆激酶 原核表达 抗体制备 纤溶活性 |
| title | Expression of nattokinase in E. coli. and preparation of its polyclonal antibody(纳豆激酶原核表达纯化及多克隆抗体的制备) |
| title_full | Expression of nattokinase in E. coli. and preparation of its polyclonal antibody(纳豆激酶原核表达纯化及多克隆抗体的制备) |
| title_fullStr | Expression of nattokinase in E. coli. and preparation of its polyclonal antibody(纳豆激酶原核表达纯化及多克隆抗体的制备) |
| title_full_unstemmed | Expression of nattokinase in E. coli. and preparation of its polyclonal antibody(纳豆激酶原核表达纯化及多克隆抗体的制备) |
| title_short | Expression of nattokinase in E. coli. and preparation of its polyclonal antibody(纳豆激酶原核表达纯化及多克隆抗体的制备) |
| title_sort | expression of nattokinase in e coli and preparation of its polyclonal antibody 纳豆激酶原核表达纯化及多克隆抗体的制备 |
| topic | 纳豆激酶 原核表达 抗体制备 纤溶活性 |
| url | https://doi.org/zjup/1008-9497.2006.33.4.447-450 |
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