牛视紫红质基因在非洲爪蟾卵母细胞中的表达：光依赖性离子电流的证明

Khorana H G, Knox B E, Nasi E, Swanson R, Thompson D A

Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.

Proc Natl Acad Sci U S A. 1988 Nov;85(21):7917-21. doi: 10.1073/pnas.85.21.7917.

Xenopus oocytes express a gene encoding bovine rhodopsin as well as its SP6 RNA polymerase-derived transcripts and total retinal mRNA. The opsin produced is in unglycosylated (30 kDa) and two glycosylated (35 kDa and 41 kDa) forms. Incubation of the cells expressing the above proteins with 11-cis-retinal generates rhodopsin, which was purified by immunoaffinity chromatography. The purified protein shows the expected UV/visible absorption spectrum and characteristic light-dependent activation of the rod outer segment GTP binding protein. Oocytes expressing rhodopsin exhibit light-dependent ionic currents that are detected by voltage-clamp techniques.

非洲爪蟾卵母细胞表达一种编码牛视紫红质的基因及其SP6 RNA聚合酶衍生的转录本和总视网膜mRNA。产生的视蛋白有未糖基化的（30 kDa）和两种糖基化的（35 kDa和41 kDa）形式。将表达上述蛋白质的细胞与11-顺式视黄醛一起孵育可产生视紫红质，通过免疫亲和层析法对其进行纯化。纯化后的蛋白质显示出预期的紫外/可见吸收光谱以及视杆外段GTP结合蛋白的特征性光依赖性激活。表达视紫红质的卵母细胞表现出光依赖性离子电流，可通过电压钳技术检测到。