Reproducibility of Fluorescent Expression from Engineered Biological Constructs in E-coli

被引：30

作者：

Beal, Jacob ^{[1
]}

Haddock-Angelli, Traci ^{[2
]}

Gershater, Markus ^{[3
]}

de Mora, Kim ^{[2
]}

Lizarazo, Meagan ^{[2
]}

Hollenhorst, Jim ^{[4
]}

Rettberg, Randy ^{[2
]}

机构：

[1] Raytheon BBN Technol, Cambridge, MA USA

[2] iGEM Fdn, Cambridge, MA USA

[3] Synthace, London, England

[4] Agilent, Santa Clara, CA USA

来源：

PLOS ONE | 2016年 / 11卷 / 03期

关键词：

CALIBRATION; STANDARD;

D O I：

10.1371/journal.pone.0150182

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页数：22

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