Abstract
Histidine decarboxylase (HDC) plays a critical role in the synthesis of histamine, a central nervous system neurotransmitter used by both vertebrates and invertebrates. Past attempts to create antisera that recognize the HDC protein in vivo have not produced satisfactory antisera. While HDC antiserum has been made in other organisms, they appear not to be useful across species, including Drosophila melanogaster. As a result, little is known about the localization as well as the biochemistry of HDC in the fly. It has been suggested that HDC undergoes a complex maturation process, including cleavage of the polypeptide at both the N- and C- termini of the predicted protein. We report an approach that should allow the HDC protein to be examined in vivo using internal epitope tagging. A plasmid containing a functional Hdc gene was modified by insertion of epitope tags, 6xHis into the protein coding region of the Hdc gene at specific sites. The location of these tags in the protein structure is predicted to be in the mature HDC protein, and thus, should be present where HDC is active. This project will allow future research investigating the biochemistry and cell biology of HDC, after germline transformation of the tagged Hdc constructs is completed. Introduction