Ginjaar H B, van Paassen H B, den Dunnen J T, Man N T, Morris G E, Moorman A F, van Ommen G J
Department of Human Genetics, Sylvius Laboratory, Leiden, The Netherlands.
FEBS Lett. 1992 Aug 24;308(3):293-7. doi: 10.1016/0014-5793(92)81296-x.
For the study of the structure and function relationship of dystrophin, defective in DMD, and for diagnostic purposes it is important to dispose of antibodies against different parts of the protein. We have made five different constructs for the expression of fusion proteins containing parts of the four domains of dystrophin. Two different recombinant expression vectors, pATH2 and pEX1, were used. Rabbits were immunized with the fusion products and several polyclonal antibodies were raised. At a later stage, monoclonal antibodies were also raised to some of the fusion proteins. One polyclonal antibody, named P20 AB, is directed against the region covering amino acid sequence 1749-2248 or the nucleotide sequence 5456-6953 of the mRNA, which corresponds to the major deletion-prone region of the DMD gene. We show the particular value, sensitivity and specificity of the P20 AB in dystrophin analysis.
为了研究杜氏肌营养不良症（DMD）中缺陷的抗肌萎缩蛋白的结构与功能关系，以及用于诊断目的，获得针对该蛋白不同部位的抗体非常重要。我们构建了五种不同的用于表达融合蛋白的构建体，这些融合蛋白包含抗肌萎缩蛋白四个结构域的部分片段。使用了两种不同的重组表达载体，pATH2和pEX1。用融合产物免疫兔子，产生了几种多克隆抗体。在后期，还针对一些融合蛋白产生了单克隆抗体。一种名为P20 AB的多克隆抗体针对覆盖mRNA氨基酸序列1749 - 2248或核苷酸序列5456 - 6953的区域，该区域对应于DMD基因的主要易缺失区域。我们展示了P20 AB在抗肌萎缩蛋白分析中的特殊价值、敏感性和特异性。
