| 粒细胞—巨噬细胞集落刺激因子／白细胞介素—3融合蛋白的表达研究 |
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| 引用本文： | 张毅,屈贤铭.粒细胞—巨噬细胞集落刺激因子／白细胞介素—3融合蛋白的表达研究[J].生物工程学报,2000,16(3):316-319. |
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| 作者姓名： | 张毅 屈贤铭 |
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| 作者单位： | 张毅（中国科学院上海生命科学院生物工程研究中心上海
200233）;屈贤铭（中国科学院上海生命科学院生物工程研究中心上海
200233）;杨胜利（中国科学院上海生命科学院生物工程研究中心上海
200233） |
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| 摘 要： | 利用ＰＣＲ扩增得到粒细胞－巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）、白细胞介素－３（ＩＬ－３）完整基因片段，将其分别克隆ｐＧＥＭ－Ｔ构建成ＧＭ－ＣＳＦ／ＩＬ－３融合蛋白基因，ＤＮＡ序列与设计预期一致。将得到的融合蛋白基因克隆对７２ＲＮＡ聚合酶表达载体ｐＴ７ｚｚ，得到表达质粒ｐＦｕ，经转化至表达宿主Ｅ．ｃｏｌｉ ＢＬ２１（ＤＥ３），在ＩＰＴＧ诱导下获得融合蛋白目的产物的直接表达。经ＳＤＳ－ＰＡＧＥ电泳鉴
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| 关 键 词： | 融合蛋白 基因表达 基因药物 GM－CSF IL－3 |
| 文章编号： | 1000-3061(2000)03-0316-04 |
| 修稿时间： | 1999年4月26日 |
Studies on the Expression of the Recombinant Human GM-CSF/IL-3 Fusion Protein |
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| ZHANG Yi\ QU Xian,Ming\ YANG Sheng,Li.Studies on the Expression of the Recombinant Human GM-CSF/IL-3 Fusion Protein[J].Chinese Journal of Biotechnology,2000,16(3):316-319. |
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| Authors: | ZHANG Yi\ QU Xian Ming\ YANG Sheng Li |
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| Institution: | Shanghai Research Center of Biotechnology, Chinese Academy of Sciences. |
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| Abstract: | A human granulocyte macrophage colony stimulating factor (GM CSF)/interleukin 3(IL 3) fusion gene with a short linker between the GM CSF and IL 3 gene has been successfully constructed and expressed in E.coli under the control of T7 promoter.The recombinant fusion protein was expressed as inclusion bodies after the IPTG induction.The yield of the GM CSF/IL 3 fusion protein was over 30% of the total cellular proteins.Western blotting results showed that the fusion protein could specifically combined with GM CSF antibody and IL 3 antibody.The biological activity was detected by the GM CSF and IL 3 dependent cell line TF 1.After solubilizing with 8mol/L urea and renaturing with dialysis against Tris.HCl solution,the refolded fusion protein showed obvious activities to maintain the growth of TF 1 cell. |
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| Keywords: | Fusion protein gene expression haemopoietic growth factors |
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