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Solubility of a luciferase-GFP fusion protein in bacterial and mammalian cells GM Nelson; Alan P Escher* Center for Molecular Biology & Gene Therapy, Loma Linda University, Loma Linda, CA 92350, USA      *(aescher@som.llu.edu) A gene construct encoding a bacterial luciferase -GFP fusion protein (MW 104 kDa) was expressed in bacterial and mammalian cells to investigate the requirements for folding in vivo of a large multidomain protein in these two cell systems. Soluble and aggregated forms of the LUX-GFP protein fusion were readily observed in Escherichia coli cells using in situ visualization of GFP and differential centrifugation followed by immunoblotting. Overexpression of the groESL chaperonin genes resulted in an increase in both luciferase and GFP activity, but with no significant increase in the amounts of soluble fusion protein relative to its aggregated form. In contrast, LUX-GFP was soluble in mammalian cells, with no detectable formation of aggregates. These results support the hypothesis that mammalian cells have the ability to fold productively large multidomain proteins with higher efficiency when compared to bacteria. [Poster: escher.alanp.52742]

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