Yue Xin-Jing, Wang Jia-Rui, Zhao Jun-Ning, Pan Zhuo, Li Yue-Zhong
State Key Laboratory of Microbial Technology, Institute of Microbial Technology, Shandong University, Qingdao, PR China.
Synth Syst Biotechnol. 2024 Apr 10;9(3):540-548. doi: 10.1016/j.synbio.2024.04.007. eCollection 2024 Sep.
The chromosomal position effect can significantly affect the transgene expression, which may provide an efficient strategy for the inauguration of alien genes in new hosts, but has been less explored rationally. The bacterium harbors a large circular high-GC genome, and the position effect in this chassis may result in a thousand-fold expression variation of alien natural products. In this study, we conducted transposon insertion at TA sites on the genome, and used enrichment and dilution indexes to respectively appraise high and low expression potentials of alien genes at insertion sites. The enrichment sites are characteristically distributed along the genome, and the dilution sites are overlapped well with the horizontal transfer genes. We experimentally demonstrated the enrichment sites as high expression integration sites (HEISs), and the dilution sites unsuitable for gene integration expression. This work highlights that HEISs are the plug-and-play sites for efficient expression of integrated genes.
染色体位置效应可显著影响转基因表达，这可能为在新宿主中引入外源基因提供一种有效策略，但尚未得到合理深入的研究。该细菌拥有一个大型环状高GC含量基因组，在这个底盘中的位置效应可能导致外源天然产物的表达变化高达千倍。在本研究中，我们在基因组的TA位点进行转座子插入，并使用富集指数和稀释指数分别评估插入位点处外源基因的高表达潜力和低表达潜力。富集位点在基因组上呈特征性分布，稀释位点与水平转移基因高度重叠。我们通过实验证明了富集位点是高表达整合位点（HEISs），而稀释位点不适合基因整合表达。这项工作突出了高表达整合位点是用于高效表达整合基因的即插即用位点。
