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采用SDS/氯仿/苯酚法和oligo(dT)纤维素亲和层析法从胚鸡脑中提取的poly(A)~+mRNA注射到非州爪蟾卵母细胞体内可以成功地被翻译为具有结合功能的GABA受体,并首次用生化方法,包括[~sH]GABA配体结合试验及药物竞争性结合试验证明了卵母细胞膜上移植GABA受体的存在。蛋白质翻译抑制剂三尖杉酯碱可以完全阻断GABA受体的翻译过程。DNA转录抑制剂放线菌素D对这一过程无影响。移植GABA受体具有受体与配体结合的饱和特性,移植GABA受体与[~sH]GABA的结合可以被调变蛋白、蝇蕈醇、双可可碱及异丙基双环磷酸酯所抑制。移植GABA受体与天然鸡脑GABA受体的生化药理学性质基本一致。
Poly (A) ~ + mRNA extracted from embryonated chicken brain by SDS / chloroform / phenol method and oligo (dT) cellulose affinity chromatography can be successfully translated into Xenopus laevis oocytes with Binding GABA receptors, and for the first time using biochemical methods, including [~ sH] GABA ligand binding assays and drug competitive binding assays to demonstrate the presence of GABA receptors on oocyte membranes. The protein translation inhibitor harringtonine completely blocks GABA receptor translation. DNA transcription inhibitor actinomycin D has no effect on this process. The transplanted GABA receptor has the saturation property of receptor binding to ligand. The binding of transplanted GABA receptor to [~ sH] GABA can be inhibited by the modified protein, muscimol, boscale and isopropyl bicyclophosphate. The biochemical and pharmacological properties of transplanted GABA receptor and natural chicken brain GABA receptor are basically the same.