Browse > Article
http://dx.doi.org/10.4014/jmb.1711.11061

Simultaneous Overexpression of Integrated Genes by Copy Number Amplification of a Mini-Yeast Artificial Chromosome  

Jung, Heo-Myung (Department of Smart Bio-Health, Dong-Eui University)
Kim, Yeon-Hee (Department of Smart Bio-Health, Dong-Eui University)
Publication Information
Journal of Microbiology and Biotechnology / v.28, no.5, 2018 , pp. 821-825 More about this Journal
Abstract
A copy number amplification system for yeast artificial chromosomes (YACs) was combined with simultaneous overexpression of genes integrated into a YAC. The chromosome VII (1,105 kb) was successfully split to 887 kb, 44 kb containing the element for copy number amplification, and a 184-kb split-YAC. The 44-kb split-mini YAC was amplified a maximum of 9-fold, and the activity of the reporter enzymes integrated into the split-mini YAC increased about 5-7-fold. These results demonstrate that the mini-YAC containing a targeted chromosome region can be readily amplified, and the specific genes in the mini-YAC could be overexpressed by increasing the copy number.
Keywords
Yeast artificial chromosome; PCR-mediated chromosome splitting; copy number amplification; simultaneous overexpression;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
연도 인용수 순위
1 Burke D, Carle GF, Olson MV. 1987. Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vector. Science 236: 806-811.   DOI
2 Pavan WJ, Hieter P, Sears D, Burkhoff A, Reeves RH. 1991. High-efficiency yeast artificial chromosome fragmentation vectors. Gene 106: 125-127.   DOI
3 Emanuel SL, Cook JR, O'Rear J, Rothstein R, Pestka S. 1995. New vectors for manipulation and selection of functional yeast artificial chromosomes (YACs) containing human DNA inserts. Gene 155: 167-174.   DOI
4 Kim YH, Kaneko Y, Fukui K, Kobayashi A, Harashima S. 2005. A yeast artificial chromosome-splitting vector designed for precise manipulation of specific plant chromosome region. J. Biosci. Bioeng. 99: 55-60.   DOI
5 Smith DR, Smyth AP, Moir DT. 1990. Amplification of large artificial chromosomes. Proc. Natl. Acad. Sci. USA 87: 8242- 8246.   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 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 ${\beta}$-xylosidase gene. J. Microbiol. Biotechnol. 8: 28-33.
10 Lee LH, Kim DY, Han MK, Oh HW, Ham SJ, Park DS, et al. 2009. Characterization of an extracellular xylanase from Bacillus sp. HY-20, a bacterium in the gut of Apis mellifera. Korean J. Microbiol. 45: 332-338.
11 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