BCR/ABL-OD结构域重组蛋白的表达和纯化  

Expression and purification of fusion protein in BCR/ABL-OD domain

作　　者：叶盛开[1] 贾洪霞[1] 林媛[2] 周鹏[3] 杨曦[3] 张萍[3] 李军锋[3] 李军林[3] 董潇[3] 张兵华[4] 梁英民[1] 

机构地区：[1]第四军医大学唐都医院血液科,陕西西安710038 [2]第四军医大学秦都口腔医学院牙体病科 [3]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 [4]解放军323医院血液科,陕西西安710054

出　　处：《第四军医大学学报》2005年第2期182-184,共3页Journal of the Fourth Military Medical University

摘　　要：目的:制备BCR/ABL OD结构域-HIS的重组蛋 白.方法:用RT-PCR方法扩增BCR/ABL OD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BL21,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.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.

关 键 词：融合蛋白质类 BCR/ABL 寡聚化结构域 重组蛋白质类 

分 类 号：R331.2[医药卫生—人体生理学]