Research Article | Open Access
Volume 2024 |Article ID 0030 |

Copper-Induced In Vivo Gene Amplification in Budding Yeast

Junyi Wang,1,4 Jingya Song,1,4 Cong Fan,1,4 Jiahao Duan,1 Kaiyuan He,1 Jifeng Yuan 1,2,3

1State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University, Fujian 361102, China
2Key Laboratory for Synthetic Biotechnology of Xiamen City, Xiamen University, Fujian 361005, China
3Shenzhen Research Institute of Xiamen University, Shenzhen 518057, China
4These authors contributed equally to this work

03 Jan 2024
29 Feb 2024
28 Mar 2024


In the biotechnological industry, multicopy gene integration represents an effective strategy to maintain a high-level production of recombinant proteins and to assemble multigene biochemical pathways. In this study, we developed copper-induced in vivo gene amplification in budding yeast for multicopy gene expressions. To make copper as an effective selection pressure, we first constructed a copper-sensitive yeast strain by deleting the CUP1 gene encoding a small metallothionein-like protein for copper resistance. Subsequently, the reporter gene fused with a proline–glutamate–serine–threonine-destabilized CUP1 was integrated at the δ sites of retrotransposon (Ty) elements to counter the copper toxicity at 100 μM Cu2+. We further demonstrated the feasibility of modulating chromosomal rearrangements for increased protein expression under higher copper concentrations. In addition, we also demonstrated a simplified design of integrating the expression cassette at the CUP1 locus to achieve tandem duplication under high concentrations of copper. Taken together, we envision that this method of copper-induced in vivo gene amplification would serve as a robust and useful method for protein overproduction and metabolic engineering applications in budding yeast.

Contact us

Lucy Wang,, +86 177 0518 5080
5 Tongwei Road, Xuanwu District, Nanjing, Jiangsu Province, China

© 2019-2023 BioDesign Research. All rights Reserved.  ISSN 2693-1257.

Back to top