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-东南大学学报医学版

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

单位：东南大学医学院,遗传学研究中心,江苏,南京,210009

分类号：Q785, Q786

出版年·卷·期（页码）：2006·25·第一期（5-8）

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Dxl6N端变体与GST的融合蛋白.

Objective To explore the expression of Dxl6 N domain in E.coli BL21(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.coli BL21(DE3).Induced by IPTG,GSTdx16N fusion protein was expressed.After rupturing E.coli BL21(DE3),GSTDxl6N was purified from the supernatant with Glutathione-sepharose4B.Result The fusion protein GST-dx16N,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.

参考文献：

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