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.

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Apr 10th, 10:00 AM Apr 10th, 11:00 AM

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.