Expression plasmid vectors with convenient subcloning sites in lambda gt11 that efficiently produce detectable tagged proteins.
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.
Takemoto
Y
Y
Institute of Immunology, Syntex-Roche, Noda, Chiba, Japan.
Sato
M
M
Furuta
M
M
Hashimoto
Y
Y
D016475:3T3 Cells; D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D000818:Animals; D017123:Bacteriophage T7 / Q000235:genetics; D010582:Bacteriophage lambda / Q000235:genetics*; D001798:Blood Proteins / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D018076:DNA, Complementary / Q000235:genetics; D016259:Genes, myc / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D006389:Hemagglutinins, Viral / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*
D048868:Adaptor Proteins, Signal Transducing; D001798:Blood Proteins; D018076:DNA, Complementary; C061909:HCLS1 protein, human; D006389:Hemagglutinins, Viral; D011993:Recombinant Fusion Proteins