Isolation of cloned ribosomal protein genes from the yeast Saccharomyces carlsbergensis.
A colony bank of yeast dna obtained by cloning HindIII-generated fragments of total yeast nuclear DNA in Escherichia coli K-12 with the vector pBR322, was screened with a radioactive RNA probe enriched for a subset of ribosomal protein mRNAs. The selected recombinant DNA molecules were hybridized with poly(A)-containing mRNA under R-loop conditions. From the DNA-RNA hybrids the respective mRNAs were melted off and translated in vitro in a rabbit reticulocyte cell-free system. The translational products were analyzed by immunoprecipitation with antibodies raised against ribosomal proteins. The identity of the ribosomal protein gene products was further established by electrophoresis on two-dimensional gels. At least 15 recombinant DNA molecules were shown to contain ribosomal protein genes. Four of them, i.e. Y65, Y89, Y113 and Y138, have been characterized preliminarily.
D004271:DNA, Fungal / Q000302:isolation & purification*; D004274:DNA, Recombinant / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005796:Genes; D009693:Nucleic Acid Hybridization; D012329:RNA, Bacterial / Q000378:metabolism; D012269:Ribosomal Proteins / Q000235:genetics*; D012440:Saccharomyces / Q000235:genetics*
D004271:DNA, Fungal; D004274:DNA, Recombinant; D012329:RNA, Bacterial; D012269:Ribosomal Proteins