Reproducibility of Fluorescent Expression from Engineered Biological Constructs in E. coli.
Reproducibility of Fluorescent Expression from Engineered Biological Constructs in E. coli.
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DOI：
10.1371/journal.pone.0150182
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
iGEM Interlab Study Contributors
中科院分区：
文献类型：
--
作者：
Beal J;Haddock-Angelli T;Gershater M;de Mora K;Lizarazo M;Hollenhorst J;Rettberg R;iGEM Interlab Study Contributors
作为2014年和2015年国际遗传工程机器（iGEM）竞赛的一部分，我们展示了在合成生物学领域进行的第一次大规模实验室间研究的结果。来自世界各地88个机构的参与者测量了E.杆菌很少有参与者能够测量绝对荧光，因此数据是根据比率进行分析的。精确度与荧光强度密切相关，范围从强启动子之间的比率的1.54倍标准偏差到最强和最弱启动子之间的比率的5.75倍，并且虽然宿主菌株不影响表达比率，但仪器的选择确实如此。这一结果表明，结果的高定量精度和再现性是可能的，同时表明需要改进实验室实践的领域。
We present results of the first large-scale interlaboratory study carried out in synthetic biology, as part of the 2014 and 2015 International Genetically Engineered Machine (iGEM) competitions. Participants at 88 institutions around the world measured fluorescence from three engineered constitutive constructs in E. coli. Few participants were able to measure absolute fluorescence, so data was analyzed in terms of ratios. Precision was strongly related to fluorescent strength, ranging from 1.54-fold standard deviation for the ratio between strong promoters to 5.75-fold for the ratio between the strongest and weakest promoter, and while host strain did not affect expression ratios, choice of instrument did. This result shows that high quantitative precision and reproducibility of results is possible, while at the same time indicating areas needing improved laboratory practices.