Establishment and application of target gene disruption system in Saccharomyces boulardii

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• 作者：Longjiang Wang (1) (2)
  Hui Sun (1) (3)
  Jie Zhang (1)
  Qing Liu (1)
  Tiantian Wang (1)
  Peipei Chen (1)
  Hongmei Li (1)
  Yihong Xiao (4)
  Fangkun Wang (1)
  Xiaomin Zhao (1) (2)
  
  1. Department of Preventive Veterinary Medicine ; College of Veterinary Medicine ; Shandong Agricultural University ; Taian City ; Shandong Province ; 271-018 ; China
  2. Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention ; Shandong Agricultural University ; Taian City ; Shandong Province ; 271-018 ; China
  3. Shandong Institute of Parasitic Diseases ; Jining City ; Shandong Province ; 272-033 ; China
  4. Department of Basic Veterinary Medicine ; College of Veterinary Medicine ; Shandong Agricultural University ; Taian City ; Shandong Province ; 271-018 ; China
  
• 关键词：target gene disruption ; Saccharomyces boulardii ; URA3 ; probiotic
• 刊名：Biotechnology and Bioprocess Engineering
• 出版年：2015
• 出版时间：February 2015
• 年：2015
• 卷：20
• 期：1
• 页码：26-36
• 全文大小：754 KB
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• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Biotechnology
  
• 出版者：The Korean Society for Biotechnology and Bioengineering
• ISSN：1976-3816

文摘

Saccharomyces boulardii is the best known probiotic yeast, widely used as a therapeutic agent for the treatment or prevention of diarrhea and intestine disorders. In the present work, we established a target gene disruption system for S. boulardii based on the Cre-loxP system used for S. cerevisiae and other fungi by screening out selection markers, working out the transformation method, and constructing essential plasmids for S. boulardii. The established system was successfully applied to the URA3 gene disruption and created an ura3 null mutant strain of S. boulardii. The system can be used for PCR mediated gene disruption, cloning mediated gene disruption, and reintroduction of the deleted gene back to the mutant. All the introduced exogenous DNAs in the gene disruption procedures were removed from the final mutant strain except the two 34 bp loxP pieces left in deleted gene loci.