| BCR/ABL-OD结构域重组蛋白的表达和纯化 |
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| 引用本文： | 叶盛开,贾洪霞,林媛,周鹏,杨曦,张萍,李军锋,李军林,董潇,张兵华,梁英民. BCR/ABL-OD结构域重组蛋白的表达和纯化[J]. 医学争鸣, 2005, 26(2): 182-184 |
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| 作者姓名： | 叶盛开 贾洪霞 林媛 周鹏 杨曦 张萍 李军锋 李军林 董潇 张兵华 梁英民 |
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| 作者单位： | 第四军医大学唐都医院血液科,陕西,西安710038;第四军医大学秦都口腔医学院牙体病科;第四军医大学基础部医学遗传学和发育生物学教研室,陕西,西安,710033;第四军医大学解放军323医院血液科,陕西,西安,710054 |
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| 摘 要： | 目的：制备BCR／ABL-0D结构域-HIS的重组蛋白．方法：用RT-PCR方法扩增BCR／ABL-0D结构域的基因片段，测序正确后将其克隆入原核表达载体pET16b中，并进行酶切鉴定，鉴定正确后转化大肠杆菌B121，构建表达His-OD融合蛋白的菌株．经IPTG诱导表达后，镍-次氮基三乙酸(Ni-NTA)琼脂糖进行亲和层析纯化，用SDS-PAGE对该融合蛋白的表达产率及蛋白纯度进行了分析．结果：获得了His-OD重组融合蛋白，纯度在95%以上，产物得率约75％．结论：成功制备高纯度His-OD融合蛋白，为进一步研究OD结构域的生物学功能奠定了基础。
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| 关 键 词： | 融合蛋白质类 BCR/ABL 寡聚化结构域 重组蛋白质类 |
| 文章编号： | 1000-2790(2005)02-0182-03 |
| 修稿时间： | 2004-05-24 |
Expression and purification of fusion protein in BCR/ABL-OD domain |
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| YE Sheng-Kai,JIA Hong-Xia,LIN Yuan,ZHOU Peng,YANG Xi,ZHANG Ping,LI Jun-Feng,LI Jun-lin,DONG Xiao,ZHANG Bing-Hua,LIANG Ying-min. Expression and purification of fusion protein in BCR/ABL-OD domain[J]. Negative, 2005, 26(2): 182-184 |
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| Authors: | YE Sheng-Kai JIA Hong-Xia LIN Yuan ZHOU Peng YANG Xi ZHANG Ping LI Jun-Feng LI Jun-lin DONG Xiao ZHANG Bing-Hua LIANG Ying-min |
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| Abstract: | AIM: To produce the BCR/ABL-OD domain fusion protein. METHODS: The cDNA encoding BCR/ABL-OD domain (oligomerization domain)was amplified by using RT-PCR method and cloned into pET16b vector. The expression plasmid was introduced into E.coli strain BL21 and His-OD fusion protein accounted for over 75% of the total bacterial protein. His-OD fusion protein was purified from the cell lysates by Ni-NTA affinity chromatography and was analyzed by SDS-PAGE gel. RESULTS: The recombinant His-OD fusion proteins were prepared with high purity (over 95%) and high recovery (about 75%). CONCLUSION: The recombinant His-OD fusion proteins will facilitate the structure-function analysis of the OD domain of BCR/ABL fusion protein. |
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| Keywords: | fusion proteins BCR/ABL oligomerization domain recombinant proteins |
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