Preparation of an antiserum using a fusion protein produced by a cDNA for rat aromatic L-amino acid decarboxylase.
Aromatic L-amino acid decarboxylase (AADC) decarboxylates L-DOPA and 5-hydroxytryptophan into dopamine and serotonin, respectively. Starting from a rat AADC cDNA clone isolated in our laboratory, we produced a beta-galactosidase-AADC fusion protein in E. coli. It was purified from inclusion bodies and injected into a rabbit. The antiserum identified AADC on a Western blot of extracts from rat organs as a unique 50 kDa band; it also strongly reacted by immunohistochemistry with dopaminergic and serotonergic neurons. This new beta-galactosidase-AADC fusion protein then constitutes a useful tool for producing AADC as an antigen free of contamination by mammalian proteins.
Biochemie cellulaire, Collège de France, Paris.
M
M
Krieger
Y
Y
Tillet
F
F
Gros
J
J
Thibault
D000818:Animals; D001142:Aromatic-L-Amino-Acid Decarboxylases / Q000276:immunology* / Q000378:metabolism; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism; D004247:DNA; D004926:Escherichia coli / Q000378:metabolism; D007106:Immune Sera / Q000276:immunology*; D007150:Immunohistochemistry; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D012680:Sensitivity and Specificity; D001616:beta-Galactosidase / Q000276:immunology
D007106:Immune Sera; D011993:Recombinant Fusion Proteins; D004247:DNA; D001616:beta-Galactosidase; D001142:Aromatic-L-Amino-Acid Decarboxylases