scholarly journals GST Fusion Protein Expression Vector for In-Frame Cloning and Site-Directed Mutagenesis

BioTechniques ◽  
1998 ◽  
Vol 24 (2) ◽  
pp. 194-196 ◽  
Author(s):  
Thérèse Hunter ◽  
Gary J. Hunter
Keyword(s):  
Fusion Protein ◽  
Protein Expression ◽  
Expression Vector ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Fusion Protein Expression ◽  
Gst Fusion Protein

Optimization of a Fusion Protein Expression System using Human Cell Lines to Create a Practical Immunoassay Reagent

2015 ◽  
Vol 41 (1) ◽  
pp. 38-42 ◽  
Author(s):  
Kounosuke Hayashi ◽  
Yusuke Tomozoe ◽  
Kenji Nagai ◽  
Kyoichi Matsuba ◽  
Masayuki Mitsumori ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Protein Expression ◽  
Cell Lines ◽  
Human Cell ◽  
Expression System ◽  
Human Cell Lines ◽  
Fusion Protein Expression ◽  
Protein Expression System

scholarly journals Mapping the BKCa Channel's “Ca2+ Bowl”

2004 ◽  
Vol 123 (5) ◽  
pp. 475-489 ◽  
Author(s):  
Lin Bao ◽  
Christina Kaldany ◽  
Ericka C. Holmstrand ◽  
Daniel H. Cox
Keyword(s):  
Fusion Protein ◽  
Binding Site ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Membrane Domain ◽  
Intracellular Domain ◽  
High Affinity ◽  
Acidic Residues ◽  
The Relationship

There is controversy over whether Ca2+ binds to the BKCa channel's intracellular domain or its integral-membrane domain and over whether or not mutations that reduce the channel's Ca2+ sensitivity act at the point of Ca2+ coordination. One region in the intracellular domain that has been implicated in Ca2+ sensing is the “Ca2+ bowl”. This region contains many acidic residues, and large Ca2+-bowl mutations eliminate Ca2+ sensing through what appears to be one type of high-affinity Ca2+-binding site. Here, through site-directed mutagenesis we have mapped the residues in the Ca2+ bowl that are most important for Ca2+ sensing. We find acidic residues, D898 and D900, to be essential, and we find them essential as well for Ca2+ binding to a fusion protein that contains a portion of the BKCa channel's intracellular domain. Thus, much of our data supports the conclusion that Ca2+ binds to the BKCa channel's intracellular domain, and they define the Ca2+ bowl's essential Ca2+-sensing motif. Overall, however, we have found that the relationship between mutations that disrupt Ca2+ sensing and those that disrupt Ca2+ binding is not as strong as we had expected, a result that raises the possibility that, when examined by gel-overlay, the Ca2+ bowl may be in a nonnative conformation.

A fusion protein expression analysis using surface plasmon resonance imaging

2004 ◽  
Vol 330 (2) ◽  
pp. 251-256 ◽  
Author(s):  
Jin-Mi Jung ◽  
Yong-Beom Shin ◽  
Min-Gon Kim ◽  
Hyeon-Su Ro ◽  
Hee-Tae Jung ◽  
...  
Keyword(s):  
Surface Plasmon Resonance ◽  
Fusion Protein ◽  
Protein Expression ◽  
Surface Plasmon ◽  
Expression Analysis ◽  
Plasmon Resonance ◽  
Surface Plasmon Resonance Imaging ◽  
Resonance Imaging ◽  
Fusion Protein Expression
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