Preparation of Protein Arrays Using Cell-Free Protein Expression

2013 ◽  
pp. 245-255 ◽  
Author(s):  
Elizabeth A. Cook ◽  
Mingyue He
Keyword(s):  
Protein Expression ◽  
Protein Arrays ◽  
Free Protein

scholarly journals Characterization of synthetic riboswitch in cell-free protein expression systems

RNA Biology ◽  
2021 ◽  
pp. 1-12
Author(s):  
Yaroslav Chushak ◽  
Svetlana Harbaugh ◽  
Kathryn Zimlich ◽  
Bryan Alfred ◽  
Jorge Chávez ◽  
...  
Keyword(s):  
Protein Expression ◽  
Expression Systems ◽  
Free Protein

scholarly journals Cell-free Protein Expression Using the Rapidly Growing Bacterium Vibrio natriegens

10.3791/59495 ◽  
2019 ◽  
Author(s):  
Daniel J. Wiegand ◽  
Henry H. Lee ◽  
Nili Ostrov ◽  
George M. Church
Keyword(s):  
Protein Expression ◽  
Free Protein

scholarly journals Expression of Recombinant Fusion Protein of Newcastle Disease Virus from Escherichia coli Plasmid Clone C-2a by In-vitro Cell-free Protein Expression System

2020 ◽  
Vol 20 ◽  
pp. 04004
Author(s):  
Ahmad Pandu Satria Wiratama ◽  
Aris Haryanto
Keyword(s):  
Protein Expression ◽  
Newcastle Disease ◽  
Recombinant Plasmid ◽  
Expression System ◽  
Free Protein ◽  
F Protein ◽  
E Coli ◽  
Protein Expression System ◽  
Sds Page

Newcastle Disease Virus (NDV) is an infectious disease that infect many kinds of wild and domesticated birds. Infection of NDV become a massive problem for poultry industry around the world especially in Indonesia. Vaccination is an effort to prevent the infection of NDV in poultry. NDV vaccine that used in Indonesia is a conventional life vaccine from LaSota and B1 strains. These type of vaccine is 21%-23% genetically distinct with the virus that spread in the environment. The antibody protection provided by the vaccine is not effective. Therefore, vaccination with new local NDV strain is needed to prevent the NDV infection in Indonesia. The previously study research reported that the local isolate of NDV from Kulon Progo, Indonesia has been isolated. Fusion (F) protein encoding gene that has been inserted into pBT7-N-His expression p lasmid which isolated from clone C-2a of E. coli, then it was expressed by the Cell-free protein expression system. The aim of this study was to confirm whether clone C-2a of E.coli carrying a recombinant plasmid pBT7-N-His-Fusion NDV and to express a recombinant F protein of NDV in-vitro from expression plasmid by cell-free protein expression system. This work started by detection of recombinant plasmid pBT7-N-His-Fusion NDV by DNA plasmid extraction followed by agarose gel electrophoresis. The recombinant F protein was in-vitro expressed by cell-free protein expression kit. The expressed F protein of NDV then was visualized by SDS-PAGE and Westernblott to analyse the expression of NDV recombinant F protein. It confirmed that clone C-2a of E. coli contained plasmid pBT7-N-His (4.001 bp) inserted by recombinant F protein of NDV gene (642 bp). The visualisation of expressed recombinant F protein by SDS-PAGE and Westernblott showed the NDV recombinant F protein was a specific protein fragment with molecular weight of 25,6 kDa..

Preparation of biomimetic gene hydrogel via polymerase chain reaction for cell-free protein expression

2019 ◽  
Vol 63 (1) ◽  
pp. 99-106 ◽  
Author(s):  
Feng Li ◽  
Wenting Yu ◽  
Xue Zhang ◽  
Xiaocui Guo ◽  
Xihan Xu ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Protein Expression ◽  
Chain Reaction ◽  
Free Protein ◽  
Polymerase Chain

A Cell-Free Protein Expression System Derived from Human Primary Peripheral Blood Mononuclear Cells

2020 ◽  
Vol 9 (8) ◽  
pp. 2188-2196
Author(s):  
David Burgenson ◽  
Jonathan Linton ◽  
Xudong Ge ◽  
Yordan Kostov ◽  
Leah Tolosa ◽  
...  
Keyword(s):  
Protein Expression ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Expression System ◽  
Peripheral Blood Mononuclear ◽  
Free Protein ◽  
Protein Expression System ◽  
Blood Mononuclear Cells ◽  
A Cell

scholarly journals Wheat germ systems for cell-free protein expression

FEBS Letters ◽  
2014 ◽  
Vol 588 (17) ◽  
pp. 2762-2773 ◽  
Author(s):  
Matthias Harbers
Keyword(s):  
Protein Expression ◽  
Wheat Germ ◽  
Free Protein

scholarly journals A novel family of expression vectors with multiple affinity tags for wheat germ cell-free protein expression

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Szilvia Krisztina Nagy ◽  
Brigitta Margit Kállai ◽  
Judit András ◽  
Tamás Mészáros
Keyword(s):  
Protein Expression ◽  
Germ Cell ◽  
Cleavage Site ◽  
Wheat Germ ◽  
In Vitro Translation ◽  
Affinity Tags ◽  
Free Protein ◽  
Target Proteins ◽  
Interaction Studies

Abstract Background Cell-free protein expression has become a widely used alternative of in vivo, cell-based systems in functional and structural studies of proteins. The wheat germ-based method outstands from the commercially available eukaryotic in vitro translation systems by its flexibility, high translation efficiency and success rate of properly folded eukaryotic protein synthesis. The original T7 promoter containing pEU3-NII vector was improved previously by addition of a ligation-independent cloning site, His6- and GST-tags, and a TEV protease cleavage site to facilitate the creation of recombinant plasmids, permit affinity purification, and enable production of purified, tag-free target proteins, respectively. Results Here, we describe a further development of pEU3-NII vector by inserting the rare-cutting, NotI restriction enzyme cleavage site to simplify vector linearization step prior to in vitro transcription. Additionally, His12, FLAG, and Halo affinity tag coding vectors have been created to increase detection sensitivity, specificity of interaction studies, and provide covalently linkable ligands for pull-down assays, respectively. Finally, the presented GST-His6, and GST-biotin double-tagging vectors could broaden the range of possibilities of protein-protein interaction studies. Conclusions The new generation of pEU3-NII vector family allows a more rapid production of translationally active mRNA and wheat germ cell-free expression of target proteins with a wide variety of affinity tags thus enables designing flexible and diverse experimental arrangement for in vitro studies of proteins.

scholarly journals Cell-Free Protein Expression under Macromolecular Crowding Conditions

PLoS ONE ◽  
2011 ◽  
Vol 6 (12) ◽  
pp. e28707 ◽  
Author(s):  
Xumeng Ge ◽  
Dan Luo ◽  
Jianfeng Xu
Keyword(s):  
Protein Expression ◽  
Macromolecular Crowding ◽  
Free Protein

Characterization of protein expression levels with label-free detected reverse phase protein arrays

2016 ◽  
Vol 509 ◽  
pp. 67-72 ◽  
Author(s):  
Xuexue Guo ◽  
Yihong Deng ◽  
Chenggang Zhu ◽  
Junlong Cai ◽  
Xiangdong Zhu ◽  
...  
Keyword(s):  
Protein Expression ◽  
Label Free ◽  
Reverse Phase ◽  
Protein Arrays ◽  
Expression Levels ◽  
Phase Protein
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