通过小型酵母人工染色体的拷贝数扩增实现整合基因的同时过表达。
Simultaneous Overexpression of Integrated Genes by Copy Number Amplification of a Mini-Yeast Artificial Chromosome.
作者信息
Jung Heo-Myung, Kim Yeon-Hee
机构信息
Department of Smart Bio-Health, Dong-Eui University, Busan 47340, Republic of Korea.
Biomedical Engineering and Biotechnology Major, Divison of Applied Bioengineering, Dong-Eui University, Busan 47340, Republic of Korea.
出版信息
J Microbiol Biotechnol. 2018 May 28;28(5):821-825. doi: 10.4014/jmb.1711.11061.
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.
用于酵母人工染色体（YAC）的拷贝数扩增系统与整合到YAC中的基因的同时过表达相结合。VII号染色体（1105 kb）成功地被分裂为887 kb、包含拷贝数扩增元件的44 kb以及一个184 kb的分裂YAC。44 kb的分裂微型YAC最多可扩增9倍，整合到分裂微型YAC中的报告酶的活性增加了约5至7倍。这些结果表明，含有目标染色体区域的微型YAC可以很容易地被扩增，并且微型YAC中的特定基因可以通过增加拷贝数而过表达。
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