Takemoto Y, Sato M, Furuta M, Hashimoto Y
Institute of Immunology, Syntex-Roche, Noda, Chiba, Japan.
DNA Cell Biol. 1997 Jul;16(7):893-6. doi: 10.1089/dna.1997.16.893.
We have generated cDNA expression vectors that efficiently produce tagged proteins. The newly introduced cloning site of this plasmid facilitates subcloning of cDNA in the lambda gt11 phage into the plasmid vector. Because the cDNA is inserted next to the motifs of the tagged DNA sequence, the protein produced by the tag sequence-coupled cDNA is easily detected by Western blot analysis or immunoprecipitation using commercially available antibodies. The double-tagged protein significantly enhances the efficiency of Western blot and immunoprecipitation detection as compared with the single-tagged protein.
我们构建了能高效产生带标签蛋白质的cDNA表达载体。该质粒新引入的克隆位点便于将λgt11噬菌体中的cDNA亚克隆到质粒载体中。由于cDNA插入到带标签DNA序列的基序旁边，通过蛋白质印迹分析或使用市售抗体进行免疫沉淀，很容易检测到由标签序列偶联的cDNA产生的蛋白质。与单标签蛋白质相比，双标签蛋白质显著提高了蛋白质印迹和免疫沉淀检测的效率。
