An improved protocol for amino acid type-selective isotope labeling in insect cells
An improved protocol for amino acid type-selective isotope labeling in insect cells
Abstract
An improved expression protocol is proposed for amino acid type-specific , -isotope labeling of proteins in baculovirus-infected (BV) insect cell cultures. This new protocol modifies the methods published by Gossert et al. (J Biomol NMR 51(4):449â??456, 2011) and provides efficient incorporation of isotopically labeled amino acids, with similar yields per L versus unlabeled expression in rich media. Gossert et al. identified the presence of unlabeled amino...
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07-15-2017 05:05 PM
An economic approach to efficient isotope labeling in insect cells using homemade 15 N-, 13 C- and 2 H-labeled yeast extracts
An economic approach to efficient isotope labeling in insect cells using homemade 15 N-, 13 C- and 2 H-labeled yeast extracts
Abstract
Heterologous expression of proteins in insect cells is frequently used for crystallographic structural studies due to the high yields even for challenging proteins requiring the eukaryotic protein processing capabilities of the host. However for NMR studies, the need for isotope labeling poses extreme challenges in eukaryotic hosts. Here, we describe a robust method to achieve uniform protein 15N and 13C labeling of up...
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06-12-2015 07:48 PM
Affordable uniform isotope labeling with 2 H, 13 C and 15 N in insect cells
Affordable uniform isotope labeling with 2 H, 13 C and 15 N in insect cells
Abstract
For a wide range of proteins of high interest, the major obstacle for NMR studies is the lack of an affordable eukaryotic expression system for isotope labeling. Here, a simple and affordable protocol is presented to produce uniform labeled proteins in the most prevalent eukaryotic expression system for structural biology, namely Spodoptera frugiperda insect cells. Incorporation levels of 80Â*% can be achieved for 15N and 13C with yields comparable to expression in...
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04-30-2015 09:13 PM
[NMR paper] Robust and low cost uniform (15)N-labeling of proteins expressed in Drosophila S2 cells and Spodoptera frugiperda Sf9 cells for NMR applications.
Robust and low cost uniform (15)N-labeling of proteins expressed in Drosophila S2 cells and Spodoptera frugiperda Sf9 cells for NMR applications.
Robust and low cost uniform (15)N-labeling of proteins expressed in Drosophila S2 cells and Spodoptera frugiperda Sf9 cells for NMR applications.
J Struct Biol. 2014 Aug 27;
Authors: Meola A, Deville C, Jeffers SA, Guardado-Calvo P, Vasiliauskaite I, Sizun C, Girard-Blanc C, Malosse C, Heijenoort CV, Chamot-Rooke J, Krey T, Guittet E, Pêtres S, Rey FA, Bontems F
Abstract
Nuclear...
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09-01-2014 07:46 PM
Robust and low cost uniform 15N-labeling of proteins expressed in Drosophila S2 cells and Spodoptera frugiperda Sf9 cells for NMR applications
Robust and low cost uniform 15N-labeling of proteins expressed in Drosophila S2 cells and Spodoptera frugiperda Sf9 cells for NMR applications
Publication date: Available online 28 August 2014
Source:Journal of Structural Biology</br>
Author(s): Annalisa Meola , Célia Deville , Scott A. Jeffers , Pablo Guardado-Calvo , Ieva Vasiliauskaite , Christina Sizun , Christine Girard-Blanc , Christian Malosse , Carine van Heijenoort , Julia Chamot-Rooke , Thomas Krey , Eric Guittet , Stéphane Pêtres , Félix A. Rey , François Bontems</br>
Nuclear magnetic...
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08-29-2014 05:36 PM
[NMR paper] High-Resolution Heteronuclear Multidimensional NMR of Proteins in Living Insect Cells Using a Baculovirus Protein Expression System.
High-Resolution Heteronuclear Multidimensional NMR of Proteins in Living Insect Cells Using a Baculovirus Protein Expression System.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--pubs.acs.org-images-pubmed-acspubs.jpg Related Articles High-Resolution Heteronuclear Multidimensional NMR of Proteins in Living Insect Cells Using a Baculovirus Protein Expression System.
J Am Chem Soc. 2013 Jan 27;
Authors: Hamatsu J, O'Donovan D, Tanaka T, Shirai T, Hourai Y, Mikawa T, Ikeya T, Mishima M, Boucher W, Smith BO, Laue ED, Shirakawa M, Ito Y
...
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02-03-2013 10:19 AM
High-Resolution HeteronuclearMultidimensional NMRof Proteins in Living Insect Cells Using a Baculovirus Protein ExpressionSystem
High-Resolution HeteronuclearMultidimensional NMRof Proteins in Living Insect Cells Using a Baculovirus Protein ExpressionSystem
Jumpei Hamatsu, Daniel O’Donovan, Takashi Tanaka, Takahiro Shirai, Yuichiro Hourai, Tsutomu Mikawa, Teppei Ikeya, Masaki Mishima, Wayne Boucher, Brian O. Smith, Ernest D. Laue, Masahiro Shirakawa and Yutaka Ito
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja310928u/aop/images/medium/ja-2012-10928u_0005.gif
Journal of the American Chemical Society
DOI: 10.1021/ja310928u...
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01-27-2013 11:41 PM
A simple protocol for amino acid type selective isotope labeling in insect cells with improved yields and high reproducibility
A simple protocol for amino acid type selective isotope labeling in insect cells with improved yields and high reproducibility
Abstract An easy to use and robust approach for amino acid type selective isotope labeling in insect cells is presented. It relies on inexpensive commercial media and can be implemented in laboratories without sophisticated infrastructure. In contrast to previous protocols, where either high protein amounts or high incorporation ratios were obtained, here we achieve both at the same time. By supplementing media with a well considered amount of yeast extract,...
Application Note: Efficient Uniform Labeling of Proteins Expressed in Baculovirus-Infected Insect Cells Using ... - SelectScience
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