Event Title
Detection of 6xHis Labeled HDC Protein in Drosophila Melanogaster
Presentation Type
Poster/Portfolio
Presenter Major(s)
Biomedical Sciences
Mentor Information
Debra Burg, Martin Burg
Department
Biomedical Sciences
Location
Kirkhof Center KC39
Start Date
10-4-2013 10:00 AM
End Date
10-4-2013 11:00 AM
Keywords
Life Science
Abstract
Histamine is a neurotransmitter used by photoreceptors in the fruit fly, Drosophila melanogaster, and is synthesized by the enzyme histidine decarboxylase (HDC). Previous studies have shown that histamine is localized to the nerve terminals where it is released as a neurotransmitter. If HDC's subcellular location and post-translational processing were better understood, potential regulatory mechanisms, which lead to histamine synthesis, could be identified. Transgenic flies bearing a functional Hdc gene with an internal 6xHIS epitope tag in a specific location were studied. Western blotting and immunocytochemical examination using a penta-HIS antibody did not provide consistent detection of the labeled 6XHIS-HDC protein. However, the site in the HDC protein used is an ideal location for epitope tagging, as it does not disrupt HDC function. Thus, a different epitope, FLAG, was inserted into the Sac1 site and flies bearing the FLAG-HDC-Sac1 transgene are currently being studied.
Detection of 6xHis Labeled HDC Protein in Drosophila Melanogaster
Kirkhof Center KC39
Histamine is a neurotransmitter used by photoreceptors in the fruit fly, Drosophila melanogaster, and is synthesized by the enzyme histidine decarboxylase (HDC). Previous studies have shown that histamine is localized to the nerve terminals where it is released as a neurotransmitter. If HDC's subcellular location and post-translational processing were better understood, potential regulatory mechanisms, which lead to histamine synthesis, could be identified. Transgenic flies bearing a functional Hdc gene with an internal 6xHIS epitope tag in a specific location were studied. Western blotting and immunocytochemical examination using a penta-HIS antibody did not provide consistent detection of the labeled 6XHIS-HDC protein. However, the site in the HDC protein used is an ideal location for epitope tagging, as it does not disrupt HDC function. Thus, a different epitope, FLAG, was inserted into the Sac1 site and flies bearing the FLAG-HDC-Sac1 transgene are currently being studied.