Molecular and Cellular Microbiology / Biomedical Sciences | Molecular Genetics, Omics, and Systems Biology
Microbiol. Biotechnol. Lett. 2019; 47(4): 667-672
https://doi.org/10.4014/mbl.1907.07004
Jung Heo-Myung and Kim Yeon-Hee *
Biomedical Engineering and Biotechnology Major, Divison of Applied Bioengineering, Dong-Eui University
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.
Keywords: Candida glabrata marker, one-step integration, Saccharomyces cerevisiae, sequential integration, xylan/xylose metabolism related genes
My Dong Lieu , Thi Thuy Hang Hoang , Huyen Nguyet Tran Nguyen and Thi Kim Thuy Dang
Microbiol. Biotechnol. Lett. 2020; 48(3): 267-275 https://doi.org/10.4014/mbl.1912.12003Kanokchan Sanoppa , Tzou-Chi Huang and Ming-Chang Wu
Microbiol. Biotechnol. Lett. 2020; 48(1): 1-11 https://doi.org/10.4014/mbl.1907.07006Jung Hye-Won and Kim Yeon-Hee
Microbiol. Biotechnol. Lett. 2019; 47(4): 662-666 https://doi.org/10.4014/mbl.1906.06008