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Transformation of Probiotic Yeast and Their Recovery from Gastrointestinal Immune Tissues Following Oral Gavage in Mice
Authors:Lauren E Hudson  Taryn P Stewart  Milo B Fasken  Anita H Corbett  Tracey J Lamb
Institution:1.Department of Pediatrics, Emory University School of Medicine;2.Department of Biochemistry, Emory University School of Medicine
Abstract:Development of recombinant oral therapy would allow for more direct targeting of the mucosal immune system and improve the ability to combat gastrointestinal disorders. Adapting probiotic yeast in particular for this approach carries several advantages. These strains have not only the potential to synthesize a wide variety of complex heterologous proteins but are also capable of surviving and protecting those proteins during transit through the intestine. Critically, however, this approach requires expertise in many diverse laboratory techniques not typically used in tandem. Furthermore, although individual protocols for yeast transformation are well characterized for commonly used laboratory strains, emphasis is placed here on alternative approaches and the importance of optimizing transformation for less well characterized probiotic strains. Detailing these methods will help facilitate discussion as to the best approaches for testing probiotic yeast as oral drug delivery vehicles and indeed serve to advance the development of this novel strategy for gastrointestinal therapy.
Keywords:Immunology  Issue 108  Saccharomyces boulardii  auxotroph  URA3  Peyer''s patch  gavage  heterologous protein  GFP  UV mutagenesis  probiotic
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