The Penn State Protein Ladder system for inexpensive protein molecular weight markers

Abstract We have created the Penn State Protein Ladder system to produce protein molecular weight markers easily and inexpensively (less than a penny a lane). The system includes plasmids which express 10, 15, 20, 30, 40, 50, 60, 80 and 100 kD proteins in E. coli. Each protein migrates appropriately...

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Autores principales:	Ryan T. Santilli, John E. Williamson, Yoshitaka Shibata, Rosalie P. Sowers, Andrew N. Fleischman, Song Tan
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4d1062aac0e3420ba2ab6b4a64e3e7ee
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spelling	oai:doaj.org-article:4d1062aac0e3420ba2ab6b4a64e3e7ee2021-12-02T15:10:39ZThe Penn State Protein Ladder system for inexpensive protein molecular weight markers10.1038/s41598-021-96051-x2045-2322https://doaj.org/article/4d1062aac0e3420ba2ab6b4a64e3e7ee2021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-96051-xhttps://doaj.org/toc/2045-2322Abstract We have created the Penn State Protein Ladder system to produce protein molecular weight markers easily and inexpensively (less than a penny a lane). The system includes plasmids which express 10, 15, 20, 30, 40, 50, 60, 80 and 100 kD proteins in E. coli. Each protein migrates appropriately on SDS-PAGE gels, is expressed at very high levels (10–50 mg per liter of culture), is easy to purify via histidine tags and can be detected directly on Western blots via engineered immunoglobulin binding domains. We have also constructed plasmids to express 150 and 250 kD proteins. For more efficient production, we have created two polycistronic expression vectors which coexpress the 10, 30, 50, 100 kD proteins or the 20, 40, 60, 80 kD proteins. 50 ml of culture is sufficient to produce 20,000 lanes of individual ladder protein or 3750 lanes of each set of coexpressed ladder proteins. These Penn State Protein Ladder expression plasmids also constitute useful reagents for teaching laboratories to demonstrate recombinant expression in E. coli and affinity protein purification, and to research laboratories desiring positive controls for recombinant protein expression and purification.Ryan T. SantilliJohn E. WilliamsonYoshitaka ShibataRosalie P. SowersAndrew N. FleischmanSong TanNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ryan T. Santilli John E. Williamson Yoshitaka Shibata Rosalie P. Sowers Andrew N. Fleischman Song Tan The Penn State Protein Ladder system for inexpensive protein molecular weight markers
description	Abstract We have created the Penn State Protein Ladder system to produce protein molecular weight markers easily and inexpensively (less than a penny a lane). The system includes plasmids which express 10, 15, 20, 30, 40, 50, 60, 80 and 100 kD proteins in E. coli. Each protein migrates appropriately on SDS-PAGE gels, is expressed at very high levels (10–50 mg per liter of culture), is easy to purify via histidine tags and can be detected directly on Western blots via engineered immunoglobulin binding domains. We have also constructed plasmids to express 150 and 250 kD proteins. For more efficient production, we have created two polycistronic expression vectors which coexpress the 10, 30, 50, 100 kD proteins or the 20, 40, 60, 80 kD proteins. 50 ml of culture is sufficient to produce 20,000 lanes of individual ladder protein or 3750 lanes of each set of coexpressed ladder proteins. These Penn State Protein Ladder expression plasmids also constitute useful reagents for teaching laboratories to demonstrate recombinant expression in E. coli and affinity protein purification, and to research laboratories desiring positive controls for recombinant protein expression and purification.
format	article
author	Ryan T. Santilli John E. Williamson Yoshitaka Shibata Rosalie P. Sowers Andrew N. Fleischman Song Tan
author_facet	Ryan T. Santilli John E. Williamson Yoshitaka Shibata Rosalie P. Sowers Andrew N. Fleischman Song Tan
author_sort	Ryan T. Santilli
title	The Penn State Protein Ladder system for inexpensive protein molecular weight markers
title_short	The Penn State Protein Ladder system for inexpensive protein molecular weight markers
title_full	The Penn State Protein Ladder system for inexpensive protein molecular weight markers
title_fullStr	The Penn State Protein Ladder system for inexpensive protein molecular weight markers
title_full_unstemmed	The Penn State Protein Ladder system for inexpensive protein molecular weight markers
title_sort	penn state protein ladder system for inexpensive protein molecular weight markers
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/4d1062aac0e3420ba2ab6b4a64e3e7ee
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The Penn State Protein Ladder system for inexpensive protein molecular weight markers

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