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为了研究果蝇中SR蛋白家族新成员Dxl6的功能,通过RT PCR得到Dxl6的全长cDNA,并根据Dxl6基因产物的功能区,分别将Dxl6中间区域(Dxl6middlepart,Dxl6MP)、Dxl6C末端RS结构域(Dxl6RSdomain,Dxl6RSD)序列亚克隆至pGEX 4T 1(His)6C及pET32a表达载体中,表达和纯化获得融合蛋白。用纯化的融合蛋白GST Dxl6RSD His和GST Dxl6MP His免疫家兔,分别得到抗Dxl6RSD和抗Dxl6MP两种抗体。WESTERNBLOT结果显示两种抗体能特异地识别在原核表达系统内表达的抗原,抗Dxl6RSD的抗体对果蝇组织中的Dxl6具有较高的特异性。
In order to study the function of Dxl6, a new member of SR protein family in Drosophila, Dxl6 full-length cDNA was obtained by RT-PCR. Dxl6middlepart (Dxl6MP), Dxl6C-terminal RS domain Dxl6RSdomain, Dxl6RSD) were subcloned into pGEX 4T 1 (His) 6C and pET32a expression vector, and expressed and purified to obtain the fusion protein. The rabbits were immunized with the purified fusion proteins GST Dxl6RSD His and GST Dxl6MP His to obtain two anti-Dxl6RSD and anti-Dxl6MP antibodies, respectively. The WESTERNBLOT results showed that the two antibodies specifically recognize the antigen expressed in the prokaryotic expression system, and the anti-Dxl6RSD antibody has higher specificity for Dxl6 in the fruit fly tissues.