Development of a genomic engineering tool in Saccharomyces cerevisiae

ASIA unversity > 醫學暨健康學院 > 食品營養與保健生技學系 > 期刊論文 > Item 310904400/79194

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题名:	Development of a genomic engineering tool in Saccharomyces cerevisiae
作者:	趙雲鵬;Chao, Yun-Peng
贡献者:	保健營養生技學系
关键词:	Genomic engineering;Recombination system;Yeast fermentation
日期:	201312
上传时间:	2014-01-06 12:20:49 (UTC+0)
摘要:	Genetic manipulation of Saccharomyces cerevisiae for production of valuable biochemicals has received considerable attentions nowadays. Plasmids are a useful tool to fulfill this goal. However, problems associated with plasmids are manifest by their instability, metabolic burden, and cell–cell variation in the copy number. This issue was addressed by developing a genetic toolbox to facilitate genomic insertion of heterologous genes and in situ fusion of an artificial promoter to genomic genes. The toolbox contains integration vectors and promoter-insertion vectors, which carry the LE-KanMX-RE cassette that comprises a KanMX module flanked by the palindromic mutant loxP sites (i.e., LE* and RE). This cassette confers on integrant strains the antibiotic-resistant trait. The inserted KanMX can be later removed by the Cre-mediated recombination between LE and RE*. As proof of principle, these vectors were utilized to integrate the XYL2 gene of Pichia stipitis into yeast genome and to place the genomic XKS1 gene under the control of the PGK1 promoter. It results in a recombinant yeast strain free of markers and plasmids. After recruitment of a heterologous XYL1 gene, the recombinant strain became capable of fermenting xylose. Our result indicates that this genetic toolbox is useful for genomic engineering of S. cerevisiae.
關聯:	Journal of the Taiwan Institute of Chemical Engineers; 45(1):24–31
显示于类别:	[食品營養與保健生技學系] 期刊論文

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