果蝇神经特异性拼接因子Dxl6变体在大肠杆菌BL21(DE3)中的表达  

Expression of Drosophila Dxl6 N-domain in E.coli BL21(DE3)

作　　者：周静[1] 袁榴娣[1] 丁洁[1] 

机构地区：[1]东南大学医学院遗传学研究中心,江苏南京210009

出　　处：《东南大学学报（医学版）》2006年第1期5-8,共4页Journal of Southeast University(Medical Science Edition)

基　　金：国家自然科学基金资助项目(30270309)

摘　　要：目的:表达与纯化果蝇中SR蛋白家族新成员Dxl6 N端变体。方法:用PCR方法扩增得到Dxl6 N端基因片段,经酶切亚克隆至pGEX-4T-1原核表达载体,在大肠杆菌BL21(DE3)中与GST融合表达,并用谷胱甘肽-sepharose4B亲和层析柱纯化融合蛋白。结果:表达产物以可溶形式经亲和层析柱纯化后获得相对分子质量约为37 000的融合蛋白。结论:成功克隆、表达并纯化了果蝇神经特异性拼接因子Dxl6 N端变体与GST的融合蛋白。Objective To explore the expression of Dxl6 N domain in E. coli BI21 （DE3） and its purification. Methods Extraction the total RNA from wild type Drosophila ORER and obtaintion the Dxl6N 270 bp gene by using RT -PCR,then Dxl6N gene was inserted into the E. coli expression vector pGEX-4T-1 （His）6C. Recombinant Dxl6N plasmid was selected and transformed into E. coil BI21 （DE3）. Induced by IPTG, GST-dxI6N fusion protein was expressed. After rupturing E. coli BI21 （DE3） , GST-DxI6N was purified from the supernatant with Glutathione-sepharose4B. Result The fusion protein GST-dxl6N,whose relative molecular weight was about 37 000 and exist half in cytoplasm in a soluble status,was purified through Glutathione-sepharose4B. Conclusion GST-Dxl6 fusion protein were obtained and lay a basis for future work.

关 键 词：Dxl6N 克隆 表达 纯化 

分 类 号：Q785[生物学—分子生物学] Q786