scholarly journals Cloning, Expression and Purification ofBacillus cereusEndochitinase in theEscherichia coliAD494(DE3)pLysS Expression System

2009 ◽  
Vol 73 (5) ◽  
pp. 1172-1174
Author(s):  
Wei-Ming CHEN ◽  
Gen-Hung CHEN ◽  
Ching-San CHEN ◽  
Shann-Tzong JIANG
Keyword(s):  
Expression System ◽  
Expression And Purification

Expression and Purification of Stable 33-kDa Soluble Human CD23 Using the Drosophila S2 Expression System

2001 ◽  
Vol 22 (2) ◽  
pp. 330-336 ◽  
Author(s):  
Sanjay S. Khandekar ◽  
Ruth J. Mayer ◽  
Donna M. Cusimano ◽  
Steven R. Katchur ◽  
Edward R. Appelbaum
Keyword(s):  
Expression System ◽  
Expression And Purification

Database Study on the Expression and Purification of Membrane Proteins

2021 ◽  
Vol 28 ◽  
Author(s):  
Chen-Yan china Zhang ◽  
Shi-Qi Zhao ◽  
Shi-Long Zhang ◽  
Li-Heng Luo ◽  
Ding-Chang Liu ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Membrane Protein ◽  
Protein Expression ◽  
Drug Targets ◽  
Expression System ◽  
Data Bank ◽  
Hek293 Cells ◽  
Expression And Purification ◽  
Beta Barrel ◽  
Membrane Protein Expression

: Membrane proteins are crucial for biological processes, and many of them are important to drug targets. Understanding the three-dimensional structures of membrane proteins are essential to evaluate their bio function and drug design. High-purity membrane proteins are important for structural determination. Membrane proteins have low yields and are difficult to purify because they tend to aggregate. We summarized membrane protein expression systems, vectors, tags, and detergents, which have deposited in the Protein Data Bank (PDB) in recent four-and-a-half years. Escherichia coli is the most expression system for membrane proteins, and HEK293 cells are the most commonly cell lines for human membrane protein expression. The most frequently vectors are pFastBac1 for alpha-helical membrane proteins, pET28a for beta-barrel membrane proteins, and pTRC99a for monotopic membrane proteins. The most used tag for membrane proteins is the 6×His-tag. FLAG commonly used for alpha-helical membrane proteins, Strep and GST for beta-barrel and monotopic membrane proteins, respectively. The detergents and their concentrations used for alpha-helical, beta-barrel, and monotopic membrane proteins are different, and DDM is commonly used for membrane protein purification. It can guide the expression and purification of membrane proteins, thus contributing to their structure and bio function studying.

scholarly journals Fast in-vitro screening of FLT3-ITD inhibitors using silkworm-baculovirus protein expression system

2021 ◽  
Author(s):  
Naoki Yamamoto ◽  
Jiro Kikuchi ◽  
Yusuke Furukawa ◽  
Naoya Shibayama
Keyword(s):  
Tyrosine Kinase ◽  
Kinase Activity ◽  
Kinase Inhibitors ◽  
Expression System ◽  
Structural Studies ◽  
Expression And Purification ◽  
Tyrosine Kinase Activity ◽  
Fast Screening ◽  
Baculovirus System

We report expression and purification of a FLT3 protein with ITD mutation (FLT3-ITD) with a steady tyrosine kinase activity using a silkworm-baculovirus system, and its application as a fast screening system of tyrosine kinase inhibitors. The FLT3-ITD protein was expressed in Bombyx mori L. pupae infected by gene-modified nucleopolyhedrovirus, and was purified as an active state. We performed an inhibition assay using 17 potential kinase inhibitors, and succeeded in identifying two potent inhibitors for FLT3-ITD. The result has paved the way for screening FLT3-ITD inhibitors in a fast and easy manner, and also for structural studies.

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