Intrinsic biocontainment: Multiplex genome safeguards combine transcriptional and recombinational control of essential yeast genes
Intrinsic biocontainment: Multiplex genome safeguards combine transcriptional and recombinational control of essential yeast genes
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DOI：
10.1073/pnas.1424704112
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发表时间：
2015-02-10
影响因子：
11.1
通讯作者：
Boeke, Jef D.
中科院分区：
文献类型：
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作者：
Cai, Yizhi;Agmon, Neta;Boeke, Jef D.
在许多情况下，如病原体工作、工程生物体的现场释放应用和知识产权保护等，都可能需要生物防护。在这里，我们描述了控制啤酒酵母，酿酒酵母的生长，使用转录和重组“保障”控制的基本基因功能。依赖于小分子存在的实际生物遏制策略要求它们在非常低的浓度下具有活性，这使得它们廉价且难以检测。组蛋白基因由诱导型启动子控制，并由30 nM雌二醇控制。工程基因的稳定性分别由位点特异性重组酶的表达调节。当两个系统都处于活动状态时，产生可行衍生物的组合频率低于检测值（
Biocontainment may be required in a wide variety of situations such as work with pathogens, field release applications of engineered organisms, and protection of intellectual properties. Here, we describe the control of growth of the brewer's yeast, Saccharomyces cerevisiae, using both transcriptional and recombinational "safeguard" control of essential gene function. Practical biocontainment strategies dependent on the presence of small molecules require them to be active at very low concentrations, rendering them inexpensive and difficult to detect. Histone genes were controlled by an inducible promoter and controlled by 30 nM estradiol. The stability of the engineered genes was separately regulated by the expression of a site-specific recombinase. The combined frequency of generating viable derivatives when both systems were active was below detection (