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Reproducibility of fluorescent expression from engineered biological constructs in E. coli

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posted on 2016-11-15, 10:17 authored by J. Beal, T. Haddock-Angelli, M. Gershater, K. De Mora, M. Lizarazo, J. Hollenhorst, R. Rettberg, P. Demling, R. Hanke, M. Osthege, A. Schechtel, S. Sudarsan, A. Zimmermann, B. Gabryelczyk, M. Ikonen, M. Salmela, M. Acar, M. F. Aktas, F. Bestepe, F. S. Ceylan, S. Cigdem, M. Dohan, M. Elitok, M. Gunduz, E. Gunduz, O. F. Hatipoglu, T. Kaya, O. Sayin, S. Tapan, O. F. Tereci, A. Uçar, M. Yilmaz, J. Barrick, A. Gutierrez, D. Mishler, J. Monk, K. Mortensen, N. Shin, E. Watkins, Y. Chen, Y. Jin, Y. Shi, H. M. Zhang, B. Ono, I. M. M. Paino, L. Ribovski, I. Silva, D. K. Zampronio, N. Birkholz, R. F. Busche, O. Konzock, S. Lippold, C. Ludwig, M. Philippi, L. Platz, C. Sigismund, S. Weber, M. Wehrs, N. Werchau, A. Wronska, Z. Z. Yen, Y. Agarwal, E. Appleton, D. Densmore, A. Esmurria, K. Lewis, A. Pacheco, M. Bruchez, D. Peters, C. Telmer, L. Wang, S. Canas-Duarte, D. Giraldo-Perez, C. Gomez-Garzon, J. Madrid-Wolff, N. Marin-Medina, V. Mazzanti, L. Rodriguez-Forero, E. Scher, R. Dowell, S. O'Hara, C. S. Pogoda, K. Shattuck, A. Altintas, A. P. Bali, R. Bech, A. Egholm, A. S. L. Hansen, K. Jensen, K. B. Karlsen, C. Mosbech, S. Belkhelfa, N. Berenger, R. Bodinier, C. Jacry
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.

Funding

Raytheon BBN Technologies, Synthace, and Agilent provided support in the form of salaries for authors JB, MG, and JH, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

History

Citation

PLoS ONE, 2016, 11(3): e0150182

Author affiliation

/Organisation/COLLEGE OF MEDICINE, BIOLOGICAL SCIENCES AND PSYCHOLOGY/MBSP Non-Medical Departments/Department of Genetics

Version

  • VoR (Version of Record)

Published in

PLoS ONE

Publisher

Public Library of Science

issn

1932-6203

eissn

1932-6203

Acceptance date

2016-02-10

Copyright date

2016

Available date

2016-11-15

Publisher version

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0150182

Language

en

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