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KMID : 0043319960190040275
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Archives of Pharmacal Research
1996 Volume.19 No. 4 p.275 ~ p.279
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Identification of Certain Sequences in the 3rd Cytoplasmic Loop of D4 Dopamine Receptor that Suppress the Bacterial Expression
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Cheong Ji-Sook
Kim Ae-Young
Kim Kyeong-Man
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Abstract
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To study the functional roles of dopamine receptors, we decided to raise antibodies against these proteins. To make antigen, we expressed the whole 3rd cytoplasmic loop of dopamine receptors in a fusion protein with glutathione-S-transferase (GST). receptors, it was successful to express and purify fusion proteins for the whole 3rd cytoplasmic loops. However, we could not express the fusion protein for the whole 3rd cytoplasmic loop of dopamine receptor in the bacteria. To study the causes that prevent the bacterial expression of the GST-fusion protein of the 3rd cytoplasmic loop of dopamine receptor, we conducted more detailed studies on dopamine receptor. To locate the region which prevents bacterial expression, we made sequential constructs in the 3rd cytoplasmic loop decreasing the size step by step, and confirmed their expressions in the SDS PAGE. It was found that certain regions of 3rd cytoplasmic loop of dopamine receptor, located in N-terminal side of the 3rd cytoplasmic loop of dopamine receptor suppress the bacterial expression of fusion protein.
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KEYWORD
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D4 dopamine receptor, GST fusion protein, Bacterial expression
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