Browse > Article
http://dx.doi.org/10.48022/mbl.2207.07008

Manipulation of Mini-Yeast Artificial Chromosome Containing Xylan Metabolism Related Genes and Mitotic Stability Analysis in Yeast  

Da-In Kang (Biomedical Engineering and Biotechnology Major, Divison of Applied Bioengineering, Dong-Eui University)
Yeon-Hee Kim (Biomedical Engineering and Biotechnology Major, Divison of Applied Bioengineering, Dong-Eui University)
Publication Information
Microbiology and Biotechnology Letters / v.50, no.3, 2022 , pp. 436-440 More about this Journal
Abstract
In this study, yeast artificial chromosome Insert (YAC) harboring genes which related xylan metabolism was constructed by using chromosome manipulation technique. For efficient chromosome manipulation, each splitting fragment (DNA module) required for splitting process was prepared and these DNA modules were transformed into Saccharomyces cerevisiae strain YKY164. By two-rounds chromosome splitting, yeast chromosome VII (1,124 kb) was split 887 kb-YAC, 45 kb-mini YAC and 198 kb-YAC and YKY183 strain containing 18 chromosomes was constructed. Splitting efficiency for chromosome manipulation was 50- 78% and expression level of foreign genes on 45 kb-mini YAC and enzyme activity were indistinguishable from that of the YKY164 strain. Furthermore, xylan-degraded products by recombinant enzymes were confirmed and mini-yeast artificial chromosome maintained stable mitotic stability without chromosome loss during 160 generations.
Keywords
Yeast artificial chromosome (YAC); PCR-mediated chromosome splitting (PCS); xylan metabolism; mitotic stability; Saccharomyces cerevisiae;
Citations & Related Records
Times Cited By KSCI : 6  (Citation Analysis)
연도 인용수 순위
1 Sasano Y, Nagasawa K, Kaboli S, Sugiyama M, Harashima S. 2016. CRISPR-PCS: a powerful new approach to inducing multiple chromosome splitting in Saccharomyces cerevisiae. Sci. Rep. 6: 30278.
2 Mitsui R, Yamada R, Ogino H. 2019. CRISPR system in the yeast Saccharomyces cerevisiae and its application in the bioproduction of useful chemicals. World J. Microbiol. Biotechnol. 35: 111.
3 Hao H, Wang X, Jia H, Yu M, Zhang X, Tang H, Zhang L. 2016. Large fragment deletion using a CRISPR/Cas9 system in Saccharomyces cerevisiae. Anal. Biochem. 509: 118-123.   DOI
4 Hassan N, Easmin F, Sasano Y, Ekino K, Taguchi H, Harashima S. 2020. Systematic approach for assessing whether undeletable chromosomal regions in Saccharomyces cerevisiae are required for cell viability. AMB Express. 10: 73.
5 Lee YG, Jin YS, Cha YL, Seo JH. 2017. Bioethanol production from cellulosic hydrolysates by engineered industrial Saccharomyces cerevisiae. Bioresour. Technol. 228: 355-361.   DOI
6 Kim YH, Sugiyama M, Yamagishi K, Kaneko Y, Fukui K, Kobayashi A, et al. 2005. A versatile and general splitting technology for generating targeted YAC subclones. Appl. Microbiol. Biotechnol. 69: 65-70.   DOI
7 Sugiyama M, Ikushima S, Nakazawa T, Kaneko Y, Harashima S. 2005. PCR-mediated repeated chromosome splitting in Saccharomyces cerevisiae. BioTechniques 38: 909-914.   DOI
8 Kim YH, Nam SW. 2010. Development of simultaneous YAC manipulation-amplification (SYMA) system by chromosome splitting technique harboring copy number amplification system. J. Life Sci. 20: 789-793.   DOI
9 Kim YH, Ishikawa D, Ha PH, Sugiyama M, Kaneko Y, Harashima S. 2006. Chromosome XII context is important for rDNA function in yeast. Nucleic Acids Res. 34: 2914-2924.   DOI
10 Park AH, Sugiyama M, Harashima S, Kim YH. 2012. Creation of an ethanol-tolerant yeast strain by genome reconstruction based on chromosome splitting technology. J. Microbiol. Biotechnol. 22: 184-189.   DOI
11 Kim MJ, Kim BH, Nam SW, Choi ES, Shin DH, Cho HY, et al. 2013. Efficient secretory expression of recombinant endoxylanase from Bacillus sp. HY-20 in Saccharomyces cerevisiae. J. Life Sci. 23: 863-868.   DOI
12 Chun YC, Jung KH, Lee JC, Park SH, Chung HK, Yoon KH. 1998. Molecular cloning and the nucleotide sequence of a Bacillus sp. KK-1 β-xylosidase gene. J. Microbiol. Biotechnol. 8: 28-33.
13 Kim SR, Kwee NR, Kim B, Jin YS. 2013. Feasibility of xylose fermentation by engineered Saccharomyces cerevisiae overexpressing endogenous aldose reductase (GRE3), xylitol dehydrogenase (XYL2), and xylulose kinase (XYL3) from Scheffersomyces stipitis. FEMS Yeast Res. 13: 312-321.   DOI
14 Lee JS, Hong SK, Lee CR, Nam SW, Jeon SJ, Kim YH. 2019. Production of ethanol (agaro-bioethanol) from agarose by unified enzymatic saccharification and fermentation in recombinant yeast. J. Microbiol. Biotechnol. 29: 625-632.   DOI
15 Jung HM, Kim YH. 2019. Comparison of methods for stable simultaneous expression of various heterologous genes in Saccharomyces cerevisiae. Microbiol. Biotechnol. Lett. 47: 667-672.   DOI