Home LiteratureArticle Details
PMID: 198749 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Application of the avidin-biotin method of gene enrichment to the isolation of long double-stranded DNA containing specific gene sequences.

Nucleic acids research ·Vol. 4 ·No. 9 ·1977-09-00 ·Pages 2961-73

Pellegrini M, Holmes DS, Manning J

Abstract

A method of enriching for long double-stranded segments of eukaryotic DNA carrying particular genes is described. A purified RNA coded for by the gene is covalently attached to biotin via the protein, cytochrome c. This modified RNA is hybridized to total nuclear, double-stranded DNA under conditions that allow the formation of R-loops. Avidin, which has a high affinity for biotin, is covalently attached to polymer spheres. The complexes of avidin-spheres with DNA:RNA-biotin R-loop hybrids band in CsCl at a much lower bouyant density than does free DNA. This density is a function of the length of DNA coupled per avidin-sphere. This method was used to prepare very long double-strands of DNA highly enriched in the coding sequences for the large rRNAs of D. melanogaster and L. donovani and the histone mRNAs of S. purpuratus.

MeSH Terms
Animals Avidin Biotin Cell Nucleus/analysis Cytochrome c Group DNA/isolation & purification,metabolism Drosophila melanogaster Genes Genetic Code Histones/biosynthesis Leishmania Methods Microscopy, Electron Nucleic Acid Conformation Nucleic Acid Hybridization RNA, Messenger/metabolism Sea Urchins
Chemicals
Cytochrome c Group Histones RNA, Messenger Avidin Biotin DNA
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Pellegrini M
Holmes D S
Manning J
References (19)
19 references, click to expand
  1. EQUILIBRIUM SEDIMENTATION OF MACROMOLECULES IN DENSITY GRADIENTS.
    Proc Natl Acad Sci U S A. 1957 Jul 15;43(7):581-8 PMID: 16590059
  2. A method for gene enrichment based on the avidin-biotin interaction. Application to the Drosophila ribosomal RNA genes.
    Biochemistry. 1977 Apr 5;16(7):1364-70 PMID: 191068
  3. Sequence arrangement of the rDNA of Drosophila melanogaster.
    Cell. 1977 Feb;10(2):213-4 PMID: 402223
  4. The structural organization of ribosomal DNA in Drosophila melanogaster.
    Cell. 1977 Feb;10(2):193-212 PMID: 402222
  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. The organization of sea urchin histone genes.
    Cell. 1975 Nov;6(3):359-69 PMID: 1052773
  7. Interspersion of repetitive and nonrepetitive DNA sequences in the Drosophila melanogaster genome.
    Cell. 1975 Feb;4(2):141-55 PMID: 804998
  8. 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
  9. Hybridization of filters with competitor DNA in the liquid phase in a standard and a micro-assay.
    Biochimie. 1974;56(9):1215-21 PMID: 4615745
  10. Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. I. Structure of F-prime factors.
    J Mol Biol. 1972 Nov 14;71(2):471-97 PMID: 4564488
  11. Buoyant density sedimentation of macromolecules in sodium iothalamate density gradients.
    J Mol Biol. 1975 Mar 5;92(3):433-48 PMID: 167175
  12. A new method of in situ hybridization.
    Chromosoma. 1975 Nov 24;53(2):107-17 PMID: 172297
  13. Determination of DNA concentration by electron microscopy.
    Anal Biochem. 1976 May 7;72:460-7 PMID: 821355
  14. Characterization of six cloned DNAs from Drosophila melanogaster, including one that contains the genes for rRNA.
    Cell. 1975 Jun;5(2):149-57 PMID: 806347
  15. Isolation of histone genes from unfractionated sea urchin DNA by subculture cloning in E. coli.
    Nature. 1975 Jun 12;255(5509):533-8 PMID: 1095932
  16. Physicochemical properties of kinetoplast DNA from Crithidia acanthocephali. Crithidia luciliae, and Trypanosoma lewisi.
    J Cell Biol. 1975 Nov;67(2PT.1):378-99 PMID: 1104639
  17. Detection of specific sequences among DNA fragments separated by gel electrophoresis.
    J Mol Biol. 1975 Nov 5;98(3):503-17 PMID: 1195397
  18. Gene linkage by RNA-DNA hybridization. I. Unique DNA sequences homologous to 4 s RNA, 5 s RNA and ribosomal RNA.
    J Mol Biol. 1968 Jun 28;34(3):661-80 PMID: 5760467
  19. Properties and composition of the isolated ribosomal DNA satellite of Xenopus laevis.
    Nature. 1968 Aug 3;219(5153):454-63 PMID: 5668424
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/4.9.2961
Published
1977-09-00
Pages
2961-73
Language
English
Region
England
NLM ID
0411011
PMCID
PMC342627
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/4.9.2961 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2