Purexpress System, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/purexpress system/product/New England Biolabs
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
1) Product Images from "A minimal biochemical route towards de novo formation of synthetic phospholipid membranes"
Article Title: A minimal biochemical route towards de novo formation of synthetic phospholipid membranes
Journal: Nature Communications
Figure Legend Snippet: Linking gene expression of FadD10 to phospholipid synthesis. a Schematic representation of the cell-free expression of FadD10 and subsequent assembly of the de novo synthesized phospholipid into vesicles in the presence of appropriate reactive precursors [TX-TL: transcription/translation]. b SDS–PAGE analysis of the expression of FadD10 in the PURExpress ® System. Lane L1: No DNA; Lane L2: DHFR DNA; Lane L3: FadD10 DNA. c HPLC/ELSD traces monitoring the formation of phospholipid 3 by incubation of PURExpress ® System with an aqueous solution of dodecanoic acid, lysolipid 2 , ATP and MgCl 2 at 37 °C in the absence (gray line) or presence (orange line) of plasmid DNA coding for FadD10. d Spinning disk confocal microscopy of the in situ formed phospholipid vesicles in the PURExpress ® System driven by FadD10 expression. Membranes were stained using 0.1 mol% Texas Red ® DHPE dye. Scale bar: 5 µm. e Localization of sfGFP-FadD10 to the membrane of the vesicles formed upon addition of the plasmid encoding the former into PURE system. External proteins were digested by Proteinase K. Scale bar: 5 µm
Techniques Used: Expressing, Synthesized, SDS Page, High Performance Liquid Chromatography, Incubation, Plasmid Preparation, Confocal Microscopy, In Situ, Staining
2) Product Images from "NusG-Dependent RNA Polymerase Pausing and Tylosin-Dependent Ribosome Stalling Are Required for Tylosin Resistance by Inducing 23S rRNA Methylation in Bacillus subtilis"
Article Title: NusG-Dependent RNA Polymerase Pausing and Tylosin-Dependent Ribosome Stalling Are Required for Tylosin Resistance by Inducing 23S rRNA Methylation in Bacillus subtilis
Figure Legend Snippet: Tylosin-dependent ribosome stalling in the leader peptide. (A) Toeprint analysis of tylosin-induced ribosome stalling during translation of the leader peptide using WT and AYA mutant templates. The toeprint (TP) identified with the WT template in the presence of tylosin (+) is marked. Sequencing lanes (A, C, G, and U) are shown. The PURExpress kit containing T7 RNAP and E. coli ribosomes was used for this analysis. (B) yxjB leader region covered by the ribosome when tylosin induces stalling. The positions of the toeprint and the ribosome peptidyl (P) and aminoacyl (A) sites are shown. Additional details are as described in the Fig. 2A legend. Experiments were performed at least twice with comparable results.
Techniques Used: Mutagenesis, Sequencing
3) Product Images from "(p)ppGpp directly regulates translation initiation during entry into quiescence"
Article Title: (p)ppGpp directly regulates translation initiation during entry into quiescence
Figure Legend Snippet: ppGpp directly inhibits translation in vitro Protein synthesis in the presence of increasing concentrations of ppGpp was measured using the PURExpress in vitro reconstituted, coupled transcription-translation system (NEB). Production of CotE-FLAG was measured via Western blot with α-FLAG (means ± SDs). n.s. p > 0.05, *p
Techniques Used: In Vitro, Western Blot
Figure Legend Snippet: IF2 is a target of ppGpp IF2 was validated in vitro as a direct target of ppGpp using IF2 mutations that reduce ppGpp binding. (A) Affinity of B. subtilis EF-G and IF2 for (p)ppGpp was compared using the differential radial capillary action of a ligand assay (DRaCALA) ( Roelofs et al., 2011 ). (means ± SDs). (B) Alignment of G1 domains of B. subtilis IF2 and EF-G. Residues in blue denote those whose chemical shifts were previously identified to be most shifted upon binding of ppGpp versus GDP. Residues in red are those that were different in EF-G versus IF2 and that were used to engineer a mutant IF2 with reduced affinity for ppGpp (G226A H230A). (C) DRaCALA-based comparison of ppGpp affinity for WT and mutant IF2 (means ± SDs). (D) in vitro sensitivity of WT and mutant IF2 was assessed using the PURExpress in vitro reconstituted, coupled transcription-translation system (NEB). WT and mutant IF2 were added at equimolar amounts to separate PURExpress reactions in the presence of 1mM ppGpp and protein synthesis was monitored by Western blot (means ± SDs). n.s. p > 0.05, *p
Techniques Used: In Vitro, Binding Assay, Mutagenesis, Western Blot
4) Product Images from "Protein Synthesis Using A Reconstituted Cell-Free System"
Article Title: Protein Synthesis Using A Reconstituted Cell-Free System
Journal: Current protocols in molecular biology / edited by Frederick M. Ausubel ... [et al.]
Figure Legend Snippet: Chromozyme cleavage assay (Roche) for a truncated version of tissue plasminogen activator protein (vtPA) synthesized in the PURExpress reactions with and without PURExpress Disulfide Bond Enhancer (PDBE). This tissue plasminogen activator contains 9 disulfide
Techniques Used: Cleavage Assay, Synthesized
Figure Legend Snippet: SDS-PAGE analysis of the reverse purification of the DHFR control protein synthesized in the PURExpress reaction. M: molecular weight standards (kDa); Lane 1: Control PURExpress reaction with no input template, Lane 2: PURExpress reaction with the DHFR
Techniques Used: SDS Page, Purification, Synthesized, Molecular Weight
Figure Legend Snippet: Scanned image of a SDS-PAGE gel of proteins synthesized in the PURExpress reactions and labeled with FluoroTect™ Green Lys . Lane 1: DHFR; lane 2: GFP; lane 3: Renilla luciferase; lane 4: Firefly luciferase; lane 5: E. coli β-galactosidase.
Techniques Used: SDS Page, Synthesized, Labeling, Luciferase
5) Product Images from "Self-assembled nanoparticle-enzyme aggregates enhance functional protein production in pure transcription-translation systems"
Article Title: Self-assembled nanoparticle-enzyme aggregates enhance functional protein production in pure transcription-translation systems
Journal: PLoS ONE
Figure Legend Snippet: Characterization of PURExpress®–QD conjugates. (A) Chemical structure of the CL4 ligand used to make the QDs colloidally stable in aqueous shown in the open dithiol configuration. (B) Agarose gel electrophoretic mobility shift assay of 523 nm emitting CdSe/CdS/ZnS core/shell/shell QDs incubated without and with a series of decreasing concentrations of the PURExpress® protein solution. Less mobility is correlated with binding to enzyme and the magnitude of this is decreased as the protein solution is serially diluted. The dashed white line indicates the location of sample wells in the gel. (C) Left—High-resolution TEM micrograph of the 523 nm emitting CdSe/CdS/ZnS core/shell/shell QDs with an average diameter of 4.1 ± 0.5 nm. A single QD is circled in red for visualization. Right—High-resolution TEM micrograph of the 625 nm emitting CdSe/ZnS core/shell QDs utilized for nanoaggregation studies due to their larger size and higher electron density which makes for easier imaging. (D) TEM micrographs of the PURExpress® protein solution (i), 625 QDs in buffer (ii), and 625 QD mixed with 0.5× PURExpress® solution at two different magnifications (iii, iv). Only when the QDs are mixed with the PURExpress® solution is clustering seen. The grey shading around the QD clusters in (iii, iv) are believed to be the PURExpress® enzymes.
Techniques Used: Agarose Gel Electrophoresis, Electrophoretic Mobility Shift Assay, Incubation, Binding Assay, Transmission Electron Microscopy, Imaging
Figure Legend Snippet: Enhancement of functional PTE production by QDs. (A) Reaction setup highlighting stopping of the CFPS reactions with kanamycin at different time points. Paraoxon hydrolysis tracked by measurement of the p -nitrophenol absorbance product. Schematic not to scale. (B) PURExpress® reaction with QDs produced functional PTE, the activity of which was monitored by absorbance. Kanamycin was added at various time points to quench translation. (C) Identical PURExpress® reaction without QDs treated in the same manner as panel (B) produced less functional PTE, resulting in less activity and p -nitrophenol product absorbance. PTE PDB ID: IPTA [ 76 ]. Other protein structures are the same as shown in Fig 1 .
Techniques Used: Functional Assay, Produced, Activity Assay
Figure Legend Snippet: sfGFP production is enhanced with QDs in diluted PURExpress® reaction conditions. (A) Production of sfGFP over time with a range of QD concentrations present versus a negative control as monitored by fluorescence. Samples were excited at 485 nm and fluorescence monitored at 510 nm [ 69 ]. Plot for all the QD concentrations can be found in S6 Fig . (B) Yield of functional sfGFP, as estimated by average fluorescence from the end-range of the reactions, over the range of QD concentrations tested (red) as compared to the QD-free reaction (grey). When tested, all samples were statistically different from the free reaction, see S7 Fig and S1 Appendix.
Techniques Used: Negative Control, Fluorescence, Functional Assay
6) Product Images from "The alarmones (p)ppGpp directly regulate translation initiation during entry into quiescence"
Article Title: The alarmones (p)ppGpp directly regulate translation initiation during entry into quiescence
Journal: Proceedings of the National Academy of Sciences of the United States of America
Figure Legend Snippet: IF2 is a target of ppGpp. IF2 was validated in vitro as a direct target of ppGpp using IF2 mutations that reduce ppGpp binding. ( A ) Affinity of B. subtilis ). (means ± SDs). ( B ) Alignment of G1 domains of B. subtilis ). Residues in red are those that differ in EF-G and IF2 and were used to engineer a mutant IF2 with reduced affinity for ppGpp (G226A H230A). ( C ) DRaCALA-based comparison of (p)ppGpp affinity for WT and mutant IF2 (means ± SDs). ( D ) In vitro sensitivity of WT and mutant IF2 was assessed using the PURExpress system (NEB). WT and mutant IF2 were added at equimolar amounts to separate PURExpress reactions in the presence of 1 mM ppGpp, and protein synthesis was monitored by Western blot (means ± SDs). ** P
Techniques Used: In Vitro, Binding Assay, Mutagenesis, Western Blot