Voith G, Dingermann T
Institute for Pharmaceutical Biology, Johann Wolfgang Goethe-University, Frankfurt/Main.
Pharmazie. 1995 Nov;50(11):758-62.
We have expressed the human m2 muscarinic receptor gene in the cellular slime mold Dictyostelium discoideum. Expression under the control of the constitutive actin 6 promoter without a D. discoideum leader peptide results in cells which seem to respond to muscarinic agonists initially, but which quickly revert to non responding cells only after a few generations. However, when expressing the hm2 gene as a fusion gene together with the CSA leader peptide under the control of the regulated D. discoideum ras promoter cells are obtained which express functional muscarinic M2 receptors in a stable manner. As expected from the typical regulation of the ras promoter, M2 receptors are expressed only during development. In ligand binding assays these heterologously expressed receptors show binding characteristics similar to authentic M2 receptors.
我们已在细胞黏菌盘基网柄菌中表达了人类M2毒蕈碱受体基因。在组成型肌动蛋白6启动子控制下表达，且没有盘基网柄菌前导肽，结果产生的细胞最初似乎对毒蕈碱激动剂有反应，但几代之后很快又恢复为无反应细胞。然而，当在受调控的盘基网柄菌ras启动子控制下，将hm2基因与CSA前导肽作为融合基因表达时，获得的细胞能稳定表达功能性毒蕈碱M2受体。正如ras启动子的典型调控所预期的那样，M2受体仅在发育过程中表达。在配体结合试验中，这些异源表达的受体显示出与天然M2受体相似的结合特性。
