Immunoscreening after hybrid selection and translation.
In this procedure, plasmid cDNA clones are screened for their ability to select a particular mRNA. Plasmid DNA is bound to nitrocellulose filters, hybridized to mRNA, washed, and the selected mRNA is eluted from the filter. Eluted mRNA is characterized by translation into 35S-labeled protein, which is identified by immunoprecipitation and denaturing (SDS) polyacrylamide gel electrophoresis. The desired clone is that which is able to select an mRNA that translates into the desired protein. This procedure can be modified to characterize cosmids and bacteriophage DNA.
Velan
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Israel Institute for Biological Research, Ness Ziona, Israel.
D004247:DNA / Q000235:genetics; D007118:Immunoassay / Q000379:methods; D007202:Indicators and Reagents; D008715:Methionine / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D012641:Selection, Genetic; D013462:Sulfur Radioisotopes; D014158:Transcription, Genetic
D007202:Indicators and Reagents; D012333:RNA, Messenger; D013462:Sulfur Radioisotopes; D004247:DNA; D008715:Methionine