Gao Qi, Zheng Xuesong
School of Perfume and Aroma Technology, Shanghai Institute of Technology, Shanghai 201418, China.
Sheng Wu Gong Cheng Xue Bao. 2013 Jun;29(6):726-34.
Constructing robust gene circuits is a fundamental work for synthetic biology. Bacteria with suicide gene circuit based on quorum-sensing will kill themselves in a controllable pattern upon certain cell density. In the media of different IPTG inducer concentration, we observed the growth and suicidal behavior of the Escherichia coli. Top10F' with such gene circuit, screened the mutants and determined their mutated loci. The results show that, with higher IPTG concentration, the more wild type bacteria were killed; as well the mutants emerged earlier and spread over the population more quickly. The sequence of plasmids in those mutants revealed that a transposon inserted into the luxR gene and therefore disrupted Quorum-Sensing of these individuals. Furthermore, the insertion sequence of the plasmid can solely result in the mutants escaping from suicide.
构建稳健的基因回路是合成生物学的一项基础工作。具有基于群体感应的自杀基因回路的细菌会在达到一定细胞密度时以可控的方式自我毁灭。在不同异丙基-β-D-硫代半乳糖苷(IPTG)诱导剂浓度的培养基中,我们观察了带有这种基因回路的大肠杆菌Top10F'的生长和自杀行为,筛选了突变体并确定了它们的突变位点。结果表明,IPTG浓度越高,被杀死的野生型细菌越多;突变体出现得越早,在群体中传播得也越快。那些突变体中质粒的序列显示,一个转座子插入到了luxR基因中,从而破坏了这些个体的群体感应。此外,质粒的插入序列能够单独导致突变体逃脱自杀。