抗大鼠γ-氨基丁酸A受体β3亚基的抗体。

Antibodies to the rat beta 3 subunit of the gamma-aminobutyric acid A receptors.

作者信息

Fernando L P, Khan Z U, De Blas A L

机构信息

Division of Molecular Biology and Biochemistry, School of Biological Sciences, University of Missouri-Kansas City 64110-2499.

出版信息

Brain Res Mol Brain Res. 1995 Jan;28(1):94-100. doi: 10.1016/0169-328x(94)00188-k.

DOI:10.1016/0169-328x(94)00188-k

PMID:7707883

Abstract

Polymerase chain reaction was used to amplify the cDNA region that codes for the large intracellular loop of the beta 3 subunit of the gamma-aminobutyric acidA/benzodiazepine receptors (GABAAR/BZDR) from rat brain. The amplified cDNA was inserted into the prokaryotic expression vector pGEX-3X and a fusion protein containing glutathione-S-transferase and beta 3 intracellular loop moieties was expressed in bacteria. The fusion protein was affinity-purified and it was used to raise a rabbit anti-beta 3 antiserum. The anti-beta 3 antiserum immunoprecipitated the gamma-aminobutyric acidA receptor from rat and bovine brain. Immunoblots of the affinity-purified GABAAR/BZDR from bovine brain revealed that the anti-beta 3 antiserum reacted with a 57 kDa peptide, whereas the monoclonal antibody 62-3G1 that recognized both beta 2 and beta 3 reacted with 55 and 57 kDa peptides. The anti-beta 3 antiserum showed specificity for the beta 3 subunit vs beta 2 and beta 1.

摘要

采用聚合酶链反应扩增大鼠脑中γ-氨基丁酸A/苯二氮䓬受体（GABAAR/BZDR）β3亚基的大细胞内环编码的cDNA区域。将扩增的cDNA插入原核表达载体pGEX-3X中，并在细菌中表达包含谷胱甘肽-S-转移酶和β3细胞内环部分的融合蛋白。对融合蛋白进行亲和纯化，并用其制备兔抗β3抗血清。抗β3抗血清免疫沉淀大鼠和牛脑中的γ-氨基丁酸A受体。对从牛脑中亲和纯化的GABAAR/BZDR进行免疫印迹分析，结果显示抗β3抗血清与一条57 kDa的肽段发生反应，而识别β2和β3的单克隆抗体62-3G1与55 kDa和57 kDa的肽段发生反应。抗β3抗血清对β3亚基相对于β2和β1表现出特异性。