Todd R D, Khurana T S, Sajovic P, Stone K R, O'Malley K L
Department of Psychiatry, Washington University School of Medicine, Saint Louis, MO 63110.
Proc Natl Acad Sci U S A. 1989 Dec;86(24):10134-8. doi: 10.1073/pnas.86.24.10134.
Using rat genomic DNA, we have established a transfected mouse fibroblast cell line that expresses a spiperone binding site with the pharmacological characteristics of a D2 dopamine receptor. The expressed D2 receptors are the product of a gene that is distinct from that reported by Bunzow et al. [Bunzow, J. R., Van Tol, H. H. M., Granoly, D. K., Albert, P., Salon, J., Christie, M., Machida, C. A., Neve, K. A. & Civelli, O. (1988) Nature (London) 336, 783-787]. Flow cytometry with the Ca2+-sensitive dye indo-1 demonstrated that activation of the expressed D2 sites resulted in increases in intracellular calcium that were dependent on the influx of external Ca2+. These general cloning procedures should be applicable to the production of cell lines expressing a variety of genes for which only functional assays are available.
利用大鼠基因组DNA，我们建立了一种转染的小鼠成纤维细胞系，该细胞系表达具有D2多巴胺受体药理学特性的螺哌隆结合位点。所表达的D2受体是一个基因的产物，该基因与Bunzow等人报道的基因不同[Bunzow, J. R., Van Tol, H. H. M., Granoly, D. K., Albert, P., Salon, J., Christie, M., Machida, C. A., Neve, K. A. & Civelli, O. (1988) Nature (London) 336, 783 - 787]。用对Ca2+敏感的染料indo - 1进行流式细胞术分析表明，所表达的D2位点的激活导致细胞内钙增加，这依赖于外部Ca2+的流入。这些通用的克隆程序应该适用于生产表达各种只有功能测定方法的基因的细胞系。
