Methods utilizing cell-free protein-synthesizing systems for the identification of recombinant DNA molecules.
Herein we outline three methods that, when coupled with cell-free protein synthesis, permit the identification of recombinant DNA molecules encoding specific polypeptides. RNAs enriched by fractionation on methylmercury hydroxide agarose gels are used to prepare sequence-specific probes. Recombinant DNA clones thus identified are further characterized as to their encoded polypeptides by either hybrid selection or hybrid arrest of translation.
D000199:Actins / Q000235:genetics; D000818:Animals; D002474:Cell-Free System; D002850:Chromatography, Gel / Q000379:methods; D004247:DNA / Q000032:analysis; D004274:DNA, Recombinant / Q000032:analysis*; D005796:Genes; D005818:Genetic Engineering / Q000379:methods; D007202:Indicators and Reagents; D007700:Kinetics; D009693:Nucleic Acid Hybridization; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics* / Q000302:isolation & purification
D000199:Actins; D004274:DNA, Recombinant; D007202:Indicators and Reagents; D012333:RNA, Messenger; D004247:DNA