| 蚯蚓纤溶酶基因P239的原核表达、纯化及活性测定 |
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| 引用本文： | 陈飞,孟宪志,李莉,单风平,吴红艳,桓明辉,陈杰.蚯蚓纤溶酶基因P239的原核表达、纯化及活性测定[J].微生物学杂志,2004,24(1):19-21. |
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| 作者姓名： | 陈飞 孟宪志 李莉 单风平 吴红艳 桓明辉 陈杰 |
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| 作者单位： | 辽宁省微生物科学研究院,辽宁,朝阳,122000 |
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| 摘 要： | 通过RT-PCR方法从蚯蚓(Lumbricus bimastus)中获得蚯蚓纤溶酶基因,命名为P-239,含有 852 个核苷酸,成熟肽编码 239 个氨基酸,通过Genbank 序列同源性比较,与已知的蚯蚓纤溶酶有一定的同源性,为首次获得的新基因.随后构建了pBV-220/P-239重组质粒,在大肠杆菌DH5α中进行原核表达,再经亲和层析柱纯化复性,其产物经活性测定,确定不仅具有激酶作用,而且具有直接溶栓活性.
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| 关 键 词： | 蚯蚓纤溶酶 基因克隆 重组表达 蛋白纯化 活性测定 |
| 文章编号： | 1005-7021(2004)01-0019-03 |
| 修稿时间： | 2003年12月1日 |
Gene Expression of Earthworm Fibrinolytic Enzymein E.coli,Purification and Bioassay |
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| CHEN Fei,MENG Xianzhi,LI Li,SHAN Fengping,WU Hongyan,HUAN Minghui,CHEN Jie.Gene Expression of Earthworm Fibrinolytic Enzymein E.coli,Purification and Bioassay[J].Journal of Microbiology,2004,24(1):19-21. |
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| Authors: | CHEN Fei MENG Xianzhi LI Li SHAN Fengping WU Hongyan HUAN Minghui CHEN Jie |
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| Abstract: | The gene of earthworm fibrinolytic enzyme from earthworm(Lumbrukinase bimastus) was obtained by RT-PCR, named P-{239},The gene consists of 852 nucleotides that code 239 amino acids. Nucleotides order compared in Genbank, share certain homology with those known Lumbrukinase,it is a new gene,P-{239} was expressed in pBV220 expression system as inclusive body protein. Then the protein was purified by G25 and Sephrose 4B.The protein shows not only fibronolytic activity,but also directly fibrous protein dissolving activity. |
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| Keywords: | Earthworm Fibrinolytic Enzyme Clong Gene Recombination Expression Purifing Protein Test Activity |
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