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Identification of Certain Sequences in the 3rd Cytoplasmic Loop of$D_4$ Dopamine Receptor that Suppress the Bacterial Expression

Cheong, Ji-Sook   (Pharmacology Laboratory, College of Pharmacy, Chonnam National UniversityUU0001112  ); Kim, Ae-Young   (Pharmacology Laboratory, College of Pharmacy, Chonnam National UniversityUU0001112  ); Kim, Kyeong-Man   (Pharmacology Laboratory, College of Pharmacy, Chonnam National UniversityUU0001112  );
  • 초록

    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). $For D_2\; and\; D_3$ 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 $D_4$ 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 $D_4$ dopamine receptor, we conducted more detailed studies on $D_4$ 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 $D_4$ dopamine receptor, located in N-terminal side of the 3rd cytoplasmic loop of $D_4$ dopamine receptor suppress the bacterial expression of fusion protein.


  • 주제어

    $D_4$ dopamine receptor .   GST fusion protein .   Bacterial expression.  

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