Isolation of a partial clone of desmoplakin-1 by antibody screening of a lambda gt11 library.
Desmoplakin-1 is a 280,000 dalton component of the intracellular portion of desmosomes. Complementary DNA (cDNA) coding for a portion of desmoplakin-1 was obtained by screening a lambda gt11 cDNA library constructed specifically for this purpose. A388 cells (a human skin squamous cell carcinoma cell line) were found to produce desmoplakin-1 and mRNA from this cell line was used to prepare a random primed cDNA library. Screening 60,000 recombinants of the library with an affinity purified anti-demoplakin-1 antibody resulted in initial identification of four clones from which one 1500 base pair clone was isolated by plaque purification. The identity of this desmoplakin clone was confirmed by demonstrating that antibody affinity purified on the protein product of the clone bound desmoplakins 1 and 2 in Western blots. Thus, the clone appears to code for the shared region of desmoplakins 1 and 2.
Feldman
S R
SR
University of North Carolina School of Medicine, Chapel Hill.
O'Keefe
E J
EJ
D003598:Cytoskeletal Proteins / Q000235:genetics* / Q000276:immunology; D004247:DNA / Q000302:isolation & purification*; D051180:Desmoplakins; D003896:Desmosomes / Q000737:chemistry*; D015723:Gene Library; D006801:Humans; D014407:Tumor Cells, Cultured
D003598:Cytoskeletal Proteins; C495294:DSP protein, human; D051180:Desmoplakins; D004247:DNA