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KMID : 1236020190470040667
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Microbiology and Biotechnology Letters
2019 Volume.47 No. 4 p.667 ~ p.672
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Comparison of Methods for Stable Simultaneous Expression of Various Heterologous Genes in Saccharomyces cerevisiae
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Jung Heo-Myung
Kim Yeon-Hee
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Abstract
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We compared two integration systems for stable expression of heterologous genes in Saccharomyces cerevisiae. A Candida glabrata-derived gene was used as the selective marker for the Cre/loxP system, and XYLP, XYLB, GRE3, and XYL2 genes were used as model heterologous genes and ligated into the universal pRSCMT vector. The resulting pRS-XylP, pRS-XylB, pRS-Gre3, and pRS-Xyl2 plasmids were sequentially integrated into yeast chromosome VII by four integration processes (marker rescue and gene integration). The four introduced genes were successfully expressed. Further, the pRS-PBG2 plasmid harboring expression cassettes for the four genes was constructed for one-step integration. The four genes that were introduced were stably maintained as a gene cluster and were simultaneously expressed. The one-step integration was more effective for the simultaneous integration and expression of the four genes related to xylan/xylose metabolism. This method will enable the generation of a useful biosystem through appropriate use of gene integration methods.
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KEYWORD
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Candida glabrata marker, one-step integration, Saccharomyces cerevisiae, sequential integration, xylan/xylose metabolism related genes
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