GENERATION OF A URACIL AUXOTROPH STRAIN OF THE PROBIOTIC YEAST SACCHAROMYCES BOULARDII AS A HOST FOR THE RECOMBINANT PROTEIN PRODUCTION

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HAMEDI HASSAN; MISAGHI ALI; MODARRESSI MOHAMMAD HOSSEIN; ZAHRAEI SALEHI TAGHI; KHORASANIZADEH DORSA; KHALAJ VAHID

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Journal: Avicenna Journal of Medical Biotechnology Year:2013 | Volume:5 | Issue:1 (16) Page(s): 29-34

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Title

GENERATION OF A URACIL AUXOTROPH STRAIN OF THE PROBIOTIC YEAST SACCHAROMYCES BOULARDII AS A HOST FOR THE RECOMBINANT PROTEIN PRODUCTION

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Keywords

PROBIOTIC

RECOMBINANT PROTEINS

URACIL

YEASTS

Abstract

Background: Saccharomyces boulardii (S.boulardii) is the best known PROBIOTIC yeast.The genetic engineering of this PROBIOTIC strain requires the availability of appropriate mutants to accept various gene constructs carrying different selection markers. As the auxotrophy selection markers are under focus, we have generated a ura3 auxotroph mutant of S. boulardii for use in further genetic manipulations.Methods: Classical UV mutagenesis was used for the generation of auxotroph mutants.The mutants were selected in the presence of 5-FOA (5-Fluoroorotic acid), URACIL and uridine. URACIL auxotrophy phenotype was confirmed by the ability of mutants to grow in the presence of URACIL and the lack of growth in the absence of this compound. To test whether the URACIL auxotrophy phenotype is due to the inactivation of URA3, the mutants were transformed with a plasmid carrying the gene. An in vitro assay was used for the analysis of acid and bile resistance capacity of these mutants Results: Three mutants were found to be ura3 auxotroph as they were able to grow only in the presence of URACIL. When the URA3 gene was added, these mutants were able to grow normally in the absence of URACIL. Further in vitro analysis showed that the acid and bile resistance capacity of one of these mutants is intact and similar to the wild type.Conclusion: A URACIL auxotroph mutant of the PROBIOTIC yeast, S.boulardii, was generated and characterized. This auxotroph strain may have potential applications in the production and delivery of the recombinant pharmacuetics into the intestinal lumen.

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APA: Copy

HAMEDI, HASSAN, MISAGHI, ALI, MODARRESSI, MOHAMMAD HOSSEIN, ZAHRAEI SALEHI, TAGHI, KHORASANIZADEH, DORSA, & KHALAJ, VAHID. (2013). GENERATION OF A URACIL AUXOTROPH STRAIN OF THE PROBIOTIC YEAST SACCHAROMYCES BOULARDII AS A HOST FOR THE RECOMBINANT PROTEIN PRODUCTION. AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB), 5(1 (16)), 29-34. SID. https://sid.ir/paper/313801/en

Vancouver: Copy

HAMEDI HASSAN, MISAGHI ALI, MODARRESSI MOHAMMAD HOSSEIN, ZAHRAEI SALEHI TAGHI, KHORASANIZADEH DORSA, KHALAJ VAHID. GENERATION OF A URACIL AUXOTROPH STRAIN OF THE PROBIOTIC YEAST SACCHAROMYCES BOULARDII AS A HOST FOR THE RECOMBINANT PROTEIN PRODUCTION. AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB)[Internet]. 2013;5(1 (16)):29-34. Available from: https://sid.ir/paper/313801/en

IEEE: Copy

HASSAN HAMEDI, ALI MISAGHI, MOHAMMAD HOSSEIN MODARRESSI, TAGHI ZAHRAEI SALEHI, DORSA KHORASANIZADEH, and VAHID KHALAJ, “GENERATION OF A URACIL AUXOTROPH STRAIN OF THE PROBIOTIC YEAST SACCHAROMYCES BOULARDII AS A HOST FOR THE RECOMBINANT PROTEIN PRODUCTION,” AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB), vol. 5, no. 1 (16), pp. 29–34, 2013, [Online]. Available: https://sid.ir/paper/313801/en

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