遗传 ›› 2004, Vol. 26 ›› Issue (4): 460-464.

• 论文 • 上一篇    下一篇

抗果蝇Dxl6蛋白抗体的制备及特异性分析

王善治;袁榴娣;万永奇;刘琍;谢 维 WANG Shan-Zhi ;YUAN Liu-Di;WAN Yong-Qi; LIU Li;XIE Wei   

  1. 东南大学遗传学研究中心,遗传与发育生物学系 南京 江苏 210009Genetic Research Center, Department of Genetics and Developmental Biology, Southeast University, Nanjing, Jiangsu, 210009, China
  • 收稿日期:1900-01-01 出版日期:2004-08-10 发布日期:2004-08-10

Preparation and Specificity Analysis of Anti-Drosophila Dxl6 Antibody

  • Received:1900-01-01 Online:2004-08-10 Published:2004-08-10

摘要:
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(Dxl6 middle part, Dxl6MP)、Dxl6 C末端RS结构域(Dxl6 RS domain, 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。WESTERN BLOT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。Abstract: In order to study the function of Dxl6 which is a novel member of SR protein family, its cDNA was cloned by RT-PCR, and the sequences of its RS domain and its middle part were subcloned into two fusion express vectors, pGEX-4T-1His(6)C and pET32a. After expressing in E.coli BL21, the truncated proteins of Dxl6 RS domain part and Dxl6 middle part in pGEX-4T-1His(6)C were purified and used to immunize rabbits. Purified antibodies against the RS domain and Dxl6 middle part were obtained by affinity chromatography with the expressed products of Dxl6 RS domain part and Dxl6 middle part in pET32a, respectively. The result shows the antibody against Dxl6 RS domain has a good specificity to Dxl6 in Drosophila larvae by Western blot analysis.

关键词: 抗体制备
Key words,
原核表达, Dxl6基因, 果蝇, SR蛋白