Analysis of choline acetyltransferase protein in temperature sensitive mutant flies using newly generated monoclonal antibody.
The protein, choline acetyltransferase (ChAT; EC 2.3.1.6), was analyzed in wild-type and two different temperature-sensitive ChAT mutants of Drosophila (Cha(ts1) and Cha(ts2)) using newly generated monoclonal antibodies. In all of the three genotypes, Western blots of crude fly head extracts showed a band stained at approximately the 80-kDa position, supporting the hypothesis that these temperature-sensitive mutants were generated by point mutation in the structural gene. The staining intensity of the bands indicated that these mutants have a lesser amount of ChAT protein than wild-type, even when they are reared at the permissive temperature (18 degrees C).
Takagawa
K
K
Division of Neurosciences, Beckman Research Institute of the City of Hope, Duarte, CA 91010, USA.
Salvaterra
P
P
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D015153:Blotting, Western; D002795:Choline O-Acetyltransferase / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D007150:Immunohistochemistry; D007700:Kinetics; D008969:Molecular Sequence Data; D009154:Mutation / Q000502:physiology*; D011233:Precipitin Tests; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013696:Temperature
D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D002795:Choline O-Acetyltransferase