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Isolation of a cloned DNA segment containing a ribosomal protein gene of Drosophila melanogaster |
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| Authors: | S Fabijanski M Pellegrini |
| Institution: | Molecular Biology Section, University of Southern California, Los Angeles, CA 90007 U.S.A. |
| Abstract: | A Drosophila genomic DNA library in the vector Charon 4 was screened using cDNA derived from the small (6S-12S) poly(A)+ mRNA of 2-6-h-old Drosophila embryos. This fraction of mRNA is enriched for ribosomal protein-coding sequences. The selected recombinants were hybridized to total mRNA under conditions which allowed for isolation of homologous mRNAs. The mRNA from these RNA/DNA hybrids was eluted and translated in vitro. The translation products were analyzed by one- and two-dimensional electrophoresis with authentic ribosomal proteins as standards. One cloned DNA segment was found to contain a ribosomal protein gene, and a sequence which hybridizes strongly to at least 5 other ribosomal protein mRNAs. |
| Keywords: | Recombinant DNA 2–6-h-old embryos electrophoresis genomic libraries AMV avian myeloblastosis virus bp base pairs DTT dithiothieitol IVT in vitro translation kb kilobase pairs NP40 Nonidet P40 PEG polyethylene glycol Pipes PVP polyvinylpyr-rolidone SDS sodium dodecyl sulfate SSC 0 15 M sodium chloride 0 015 M sodium citrate |
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