Gemini, a bifunctional enzymatic and fluorescent reporter of gene expression.

<h4>Background</h4>The development of collections of quantitatively characterized standard biological parts should facilitate the engineering of increasingly complex and novel biological systems. The existing enzymatic and fluorescent reporters that are used to characterize biological pa...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Lance Martin, Austin Che, Drew Endy
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2009
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e00b98f2f6d344c99f5342760587efe0
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:e00b98f2f6d344c99f5342760587efe0
record_format	dspace
spelling	oai:doaj.org-article:e00b98f2f6d344c99f5342760587efe02021-11-25T06:28:20ZGemini, a bifunctional enzymatic and fluorescent reporter of gene expression.1932-620310.1371/journal.pone.0007569https://doaj.org/article/e00b98f2f6d344c99f5342760587efe02009-11-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19888458/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>The development of collections of quantitatively characterized standard biological parts should facilitate the engineering of increasingly complex and novel biological systems. The existing enzymatic and fluorescent reporters that are used to characterize biological part functions exhibit strengths and limitations. Combining both enzymatic and fluorescence activities within a single reporter protein would provide a useful tool for biological part characterization.<h4>Methodology/principal findings</h4>Here, we describe the construction and quantitative characterization of Gemini, a fusion between the beta-galactosidase (beta-gal) alpha-fragment and the N-terminus of full-length green fluorescent protein (GFP). We show that Gemini exhibits functional beta-gal activity, which we assay with plates and fluorometry, and functional GFP activity, which we assay with fluorometry and microscopy. We show that the protein fusion increases the sensitivity of beta-gal activity and decreases the sensitivity of GFP.<h4>Conclusions/significance</h4>Gemini is therefore a bifunctional reporter with a wider dynamic range than the beta-gal alpha-fragment or GFP alone. Gemini enables the characterization of gene expression, screening assays via enzymatic activity, and quantitative single-cell microscopy or FACS via fluorescence activity. The analytical flexibility afforded by Gemini will likely increase the efficiency of research, particularly for screening and characterization of libraries of standard biological parts.Lance MartinAustin CheDrew EndyPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 4, Iss 11, p e7569 (2009)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Lance Martin Austin Che Drew Endy Gemini, a bifunctional enzymatic and fluorescent reporter of gene expression.
description	<h4>Background</h4>The development of collections of quantitatively characterized standard biological parts should facilitate the engineering of increasingly complex and novel biological systems. The existing enzymatic and fluorescent reporters that are used to characterize biological part functions exhibit strengths and limitations. Combining both enzymatic and fluorescence activities within a single reporter protein would provide a useful tool for biological part characterization.<h4>Methodology/principal findings</h4>Here, we describe the construction and quantitative characterization of Gemini, a fusion between the beta-galactosidase (beta-gal) alpha-fragment and the N-terminus of full-length green fluorescent protein (GFP). We show that Gemini exhibits functional beta-gal activity, which we assay with plates and fluorometry, and functional GFP activity, which we assay with fluorometry and microscopy. We show that the protein fusion increases the sensitivity of beta-gal activity and decreases the sensitivity of GFP.<h4>Conclusions/significance</h4>Gemini is therefore a bifunctional reporter with a wider dynamic range than the beta-gal alpha-fragment or GFP alone. Gemini enables the characterization of gene expression, screening assays via enzymatic activity, and quantitative single-cell microscopy or FACS via fluorescence activity. The analytical flexibility afforded by Gemini will likely increase the efficiency of research, particularly for screening and characterization of libraries of standard biological parts.
format	article
author	Lance Martin Austin Che Drew Endy
author_facet	Lance Martin Austin Che Drew Endy
author_sort	Lance Martin
title	Gemini, a bifunctional enzymatic and fluorescent reporter of gene expression.
title_short	Gemini, a bifunctional enzymatic and fluorescent reporter of gene expression.
title_full	Gemini, a bifunctional enzymatic and fluorescent reporter of gene expression.
title_fullStr	Gemini, a bifunctional enzymatic and fluorescent reporter of gene expression.
title_full_unstemmed	Gemini, a bifunctional enzymatic and fluorescent reporter of gene expression.
title_sort	gemini, a bifunctional enzymatic and fluorescent reporter of gene expression.
publisher	Public Library of Science (PLoS)
publishDate	2009
url	https://doaj.org/article/e00b98f2f6d344c99f5342760587efe0
work_keys_str_mv	AT lancemartin geminiabifunctionalenzymaticandfluorescentreporterofgeneexpression AT austinche geminiabifunctionalenzymaticandfluorescentreporterofgeneexpression AT drewendy geminiabifunctionalenzymaticandfluorescentreporterofgeneexpression
_version_	1718413713728339968

Gemini, a bifunctional enzymatic and fluorescent reporter of gene expression.

Ejemplares similares