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GDNF-Luciferase fusion is fully functional upon secretion from mammalian cells Hua H Liu*; A Szalay Dept Biochemistry, Dept Pharmacology/Physiology, 11085 Campus Street, Loma Linda, CA 92350, USA      *(liu.howard@mailandnews.com) We have developed a reporter system to monitor the presence of glial cell-line derived neurotrophic factor (GDNF) in body fluids. The reporter gene is a mutant form of Renilla Luciferase cDNA which was fused with GDNF cDNA via a linker to generate a single GDNF-RUC fusion protein. Upon transfection of mammalian cells with the fusion construct, luciferase activity was detected in the cells as well as in the culture medium. The fusion of GDNF was confirmed by the fetal sheep dorsal root ganglion assay which observed distinct stimulation of neurite outgrowth in vitro. The molecular weight of the fusion is 65 KD as determined by immunoblot assay using both antibodies against Renilla Luciferase and GDNF. The amount of GDNF was determined by ELISA assay suggesting the one to one ratio of Renilla Luciferase and GDNF in the fusion protein. In addition, the expression of the fusion protein in different mammalian cell lines followed by implantation of the transformed mammalian cells into model animals will be presented. Lastly, the potential applications of the fusion protein in cellular implantation imaging and gene therapy will be discussed. [Poster: liu.huah.22022]

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