Construction of dystrophin fusion proteins to raise targeted antibodies to different epitopes

FEBS Letters

Moorman, A.F.M.¹  van Paassen, H.B.M.³  van Ommen, G.J.B.³  Morris, G.E.²  Ginjaar, H.B.³  den Dunnen, J.T.³  thi Man, Nguyen² 

[1] Department of Anatomy and Embryology, AMC, 1105 AZ Amsterdam, The Netherlands;Research Division, N.E. Wales Institute, Deeside, Clwyd, CH5 4BR, UK;Department of Human Genetics, Sylvius Laboratory, 333 AL Leiden, The Netherlands

关键词: Dystrophin;    pATH2 fusion protein;    pEX1 fusion protein;    Duchenne muscular dystrophy: P20 mutation hotspot;    Polyclenal antiserum;   

DOI  :  10.1016/0014-5793(92)81296-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

For the study of the structure and function relationship of dystrophin, defective in DMD, and for diagnostic purposes it is important to dispose of untibodies 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.

【 授权许可】

Unknown