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PMID: 379820 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The use of R-looping for structural gene identification and mRNA purification.

Nucleic acids research ·Vol. 6 ·No. 7 ·1979-06-11 ·Pages 2483-97

Woolford JL, Rosbash M

Abstract

A method is presented for the purification of mRNAs and the identification of structural gene sequences in recombinant DNA molecules. RNA is hybridized to double-stranded linear DNA such that R-loops are formed between most DNAs and their complementary RNA sequences. These R-loops are purified from unhybridized RNAs by gel filtration chromatography in the presence of a high concentration of salt. The complementary RNAs are released from the R-loops by heating, and are assayed by gel electrophoresis or cell free translation to determine their purity and to identify the proteins for which they code. We have demonstrated that recombinant DNAs containing sequences for abundant or moderately abundant mRNAs of Saccharomyces cerevisiae can be identified by this means.

MeSH Terms
Base Sequence DNA, Recombinant Escherichia coli/analysis Genes Nucleic Acid Conformation Nucleic Acid Hybridization Plasmids RNA, Messenger/isolation & purification Saccharomyces cerevisiae/analysis
Chemicals
DNA, Recombinant RNA, Messenger
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Woolford J L
Rosbash M
References (24)
24 references, click to expand
  1. SEDIMENTATION STUDIES OF THE SIZE AND SHAPE OF DNA.
    J Mol Biol. 1965 Feb;11:373-90 PMID: 14290352
  2. Analysis of mRNA populations by cDNA.mRNA hybrid-mediated inhibition of cell-free protein synthesis.
    Proc Natl Acad Sci U S A. 1978 Mar;75(3):1217-21 PMID: 274713
  3. Rates of formation and thermal stabilities of RNA:DNA and DNA:DNA duplexes at high concentrations of formamide.
    Nucleic Acids Res. 1977;4(5):1539-52 PMID: 19730
  4. Messenger RNA in heat-shocked Drosophila cells.
    J Mol Biol. 1977 Feb 5;109(4):559-87 PMID: 403287
  5. R loop mapping of the 18S and 28S sequences in the long and short repeating units of Drosophila melanogaster rDNA.
    Cell. 1977 Feb;10(2):177-92 PMID: 402221
  6. Isolation and analysis of recombinant DNA molecules containing yeast DNA.
    Gene. 1978 Sep;4(1):37-49 PMID: 365691
  7. Functional expression of cloned yeast DNA in Escherichia coli.
    Proc Natl Acad Sci U S A. 1977 Feb;74(2):487-91 PMID: 322128
  8. Regulation of a set of abundant mRNA sequences.
    Cell. 1977 Mar;10(3):463-7 PMID: 321130
  9. Positions of sea urchin (Strongylocentrotus purpuratus) histone genes relative to restriction endonuclease sites on the chimeric plasmids pSp2 and pSp17.
    Biochemistry. 1977 Apr 5;16(7):1504-12 PMID: 321020
  10. Spliced segments at the 5' terminus of adenovirus 2 late mRNA.
    Proc Natl Acad Sci U S A. 1977 Aug;74(8):3171-5 PMID: 269380
  11. Structural gene identification and mapping by DNA-mRNA hybrid-arrested cell-free translation.
    Proc Natl Acad Sci U S A. 1977 Oct;74(10):4370-4 PMID: 270678
  12. An amazing sequence arrangement at the 5' ends of adenovirus 2 messenger RNA.
    Cell. 1977 Sep;12(1):1-8 PMID: 902310
  13. Hybridization of RNA to double-stranded DNA: formation of R-loops.
    Proc Natl Acad Sci U S A. 1976 Jul;73(7):2294-8 PMID: 781674
  14. Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cell-free system from commercial wheat germ.
    Proc Natl Acad Sci U S A. 1973 Aug;70(8):2330-4 PMID: 4525168
  15. Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis.
    Biochemistry. 1973 Jul 31;12(16):3055-63 PMID: 4354250
  16. A system for mapping DNA sequences in the chromosomes of Drosophila melanogaster.
    Cell. 1974 Dec;3(4):315-25 PMID: 4216403
  17. Isolation of high molecular weight DNA from yeast nuclei.
    Anal Biochem. 1972 Sep;49(1):276-84 PMID: 4562899
  18. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels.
    Eur J Biochem. 1974 Jul 1;46(1):83-8 PMID: 4850204
  19. A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane.
    J Mol Biol. 1965 Jul;12(3):829-42 PMID: 4955314
  20. Genetic assay for small fragments of bacteriophage phi X174 deoxyribonucleic acid.
    J Virol. 1971 Aug;8(2):181-9 PMID: 4940243
  21. Coordinate regulation of the synthesis of eukaryotic ribosomal proteins.
    Proc Natl Acad Sci U S A. 1976 May;73(5):1547-51 PMID: 775493
  22. Functional genetic expression of eukaryotic DNA in Escherichia coli.
    Proc Natl Acad Sci U S A. 1976 May;73(5):1471-5 PMID: 775490
  23. Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis.
    Biochemistry. 1975 Aug 26;14(17):3787-94 PMID: 1174504
  24. Primary structure of wheat germ phenylalanine transfer RNA.
    Proc Natl Acad Sci U S A. 1969 Mar;62(3):941-5 PMID: 5257014
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/6.7.2483
Published
1979-06-11
Pages
2483-97
Language
English
Region
England
NLM ID
0411011
PMCID
PMC327867
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/6.7.2483 PMC Full Text View journal details
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