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KMID : 0545120180280050821
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Journal of Microbiology and Biotechnology
2018 Volume.28 No. 5 p.821 ~ p.825
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Simultaneous Overexpression of Integrated Genes by Copy Number Amplification of a Mini-Yeast Artificial Chromosome
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Jung Heo-Myung
Kim Yeon-Hee
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Abstract
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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.
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KEYWORD
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Yeast artificial chromosome (YAC), PCR-mediated chromosome splitting (PCS), copy number amplification, simultaneous overexpression
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