High-throughput expression of fusion proteins.
The expression of putative ORFs as fusion proteins can accelerate research greatly. The availability of an epitope tag allows the use of Western blotting as an efficient means to identify useful recombinant plasmids, which can then be used to study protein function. In addition, the epitope tag can be extremely useful in downstream applications such as protein purification, immunolocalization, and immunoprecipitation experiments. The preceding protocols should be applicable to a variety of expression vectors, and should be useful in the identification of functional plasmids. The protocols require no exotic equipment and can be adapted for use in high-throughput transfection assays, protein purification protocols, and immunolocalization studies.
Nasoff
M
M
Genomics Institute of the Novartis Research Foundation, San Diego, California 92121-1125, USA.
Bergseid
M
M
Hoeffler
J P
JP
Heyman
J A
JA
D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D008322:Mammals; D016366:Open Reading Frames; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D013879:Thioredoxins / Q000235:genetics; D014158:Transcription, Genetic; D014162:Transfection / Q000379:methods
D011993:Recombinant Fusion Proteins; D013879:Thioredoxins