Europe PMC

This website requires cookies, and the limited processing of your personal data in order to function. By using the site you are agreeing to this as outlined in our privacy notice and cookie policy.

Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA.

Author information

Affiliations

  • 1. Institut für Medizinische Mikrobiologie, Medizinische Hochschule Hannover, FRG.
      Authors
    • Edwards U 1
    (1 author)

Nucleic Acids Research, 01 Oct 1989, 17(19):7843-7853
https://doi.org/10.1093/nar/17.19.7843 PMID: 2798131 PMCID: PMC334891

Free full text in Europe PMC

Abstract 

Using a set of synthetic oligonucleotides homologous to broadly conserved sequences in-vitro amplification via the polymerase chain reaction followed by direct sequencing results in almost complete nucleotide determination of a gene coding for 16S ribosomal RNA. As a model system the nucleotide sequence of the 16S rRNA gene of M.kansasii was determined and found to be 98.7% homologous to that of M.bovis BCG. This is the first report on a contiguous sequence information of an entire amplified gene spanning 1.5 kb without any subcloning procedures.

Free full text 

Logo of narLink to Publisher's site
Nucleic Acids Res. 1989 Oct 11; 17(19): 7843–7853.
PMCID: PMC334891
PMID: 2798131

Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA.

U Edwards, T Rogall, H Blöcker, M Emde, and E C Böttger
Author information Copyright and License information Disclaimer

Abstract

Using a set of synthetic oligonucleotides homologous to broadly conserved sequences in-vitro amplification via the polymerase chain reaction followed by direct sequencing results in almost complete nucleotide determination of a gene coding for 16S ribosomal RNA. As a model system the nucleotide sequence of the 16S rRNA gene of M.kansasii was determined and found to be 98.7% homologous to that of M.bovis BCG. This is the first report on a contiguous sequence information of an entire amplified gene spanning 1.5 kb without any subcloning procedures.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (930K), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.
 
icon of scanned page 7843
7843
icon of scanned page 7844
7844
icon of scanned page 7845
7845
icon of scanned page 7846
7846
icon of scanned page 7847
7847
icon of scanned page 7848
7848
icon of scanned page 7849
7849
icon of scanned page 7850
7850
icon of scanned page 7851
7851
icon of scanned page 7852
7852
icon of scanned page 7853
7853
 

Images in this article

Image
Image
on p.7848
Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Saiki RK, Scharf S, Faloona F, Mullis KB, Horn GT, Erlich HA, Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. [Abstract] [Google Scholar]
  • Mullis KB, Faloona FA. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987;155:335–350. [Abstract] [Google Scholar]
  • Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. [Abstract] [Google Scholar]
  • Wong C, Dowling CE, Saiki RK, Higuchi RG, Erlich HA, Kazazian HH., Jr Characterization of beta-thalassaemia mutations using direct genomic sequencing of amplified single copy DNA. Nature. 330(6146):384–386. [Abstract] [Google Scholar]
  • Wrischnik LA, Higuchi RG, Stoneking M, Erlich HA, Arnheim N, Wilson AC. Length mutations in human mitochondrial DNA: direct sequencing of enzymatically amplified DNA. Nucleic Acids Res. 1987 Jan 26;15(2):529–542. [Europe PMC free article] [Abstract] [Google Scholar]
  • Gyllensten UB, Erlich HA. Generation of single-stranded DNA by the polymerase chain reaction and its application to direct sequencing of the HLA-DQA locus. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7652–7656. [Europe PMC free article] [Abstract] [Google Scholar]
  • Innis MA, Myambo KB, Gelfand DH, Brow MA. DNA sequencing with Thermus aquaticus DNA polymerase and direct sequencing of polymerase chain reaction-amplified DNA. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9436–9440. [Europe PMC free article] [Abstract] [Google Scholar]
  • Nakamaye KL, Gish G, Eckstein F, Vosberg HP. Direct sequencing of polymerase chain reaction amplified DNA fragments through the incorporation of deoxynucleoside alpha-thiotriphosphates. Nucleic Acids Res. 1988 Nov 11;16(21):9947–9959. [Europe PMC free article] [Abstract] [Google Scholar]
  • Higuchi R, Krummel B, Saiki RK. A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions. Nucleic Acids Res. 1988 Aug 11;16(15):7351–7367. [Europe PMC free article] [Abstract] [Google Scholar]
  • Scharf SJ, Horn GT, Erlich HA. Direct cloning and sequence analysis of enzymatically amplified genomic sequences. Science. 1986 Sep 5;233(4768):1076–1078. [Abstract] [Google Scholar]
  • Todd JA, Bell JI, McDevitt HO. HLA-DQ beta gene contributes to susceptibility and resistance to insulin-dependent diabetes mellitus. Nature. 1987 Oct 15;329(6140):599–604. [Abstract] [Google Scholar]
  • Dunning AM, Talmud P, Humphries SE. Errors in the polymerase chain reaction. Nucleic Acids Res. 1988 Nov 11;16(21):10393–10393. [Europe PMC free article] [Abstract] [Google Scholar]
  • Päbo S, Wilson AC. Polymerase chain reaction reveals cloning artefacts. Nature. 1988 Aug 4;334(6181):387–388. [Abstract] [Google Scholar]
  • Gray MW, Sankoff D, Cedergren RJ. On the evolutionary descent of organisms and organelles: a global phylogeny based on a highly conserved structural core in small subunit ribosomal RNA. Nucleic Acids Res. 1984 Jul 25;12(14):5837–5852. [Europe PMC free article] [Abstract] [Google Scholar]
  • Dams E, Hendriks L, Van de Peer Y, Neefs JM, Smits G, Vandenbempt I, De Wachter R. Compilation of small ribosomal subunit RNA sequences. Nucleic Acids Res. 1988;16 (Suppl):r87–173. [Europe PMC free article] [Abstract] [Google Scholar]
  • Woese CR. Bacterial evolution. Microbiol Rev. 1987 Jun;51(2):221–271. [Europe PMC free article] [Abstract] [Google Scholar]
  • Lane DJ, Pace B, Olsen GJ, Stahl DA, Sogin ML, Pace NR. Rapid determination of 16S ribosomal RNA sequences for phylogenetic analyses. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6955–6959. [Europe PMC free article] [Abstract] [Google Scholar]
  • Suzuki Y, Nagata A, Ono Y, Yamada T. Complete nucleotide sequence of the 16S rRNA gene of Mycobacterium bovis BCG. J Bacteriol. 1988 Jun;170(6):2886–2889. [Europe PMC free article] [Abstract] [Google Scholar]
  • Johnson AM, Baverstock PR. Rapid ribosomal RNA sequencing and the phylogenetic analysis of protists. Parasitol Today. 1989 Apr;5(4):102–105. [Abstract] [Google Scholar]
  • Göbel U, Maas R, Haun G, Vinga-Martins C, Stanbridge EJ. Synthetic oligonucleotide probes complementary to rRNA for group- and species-specific detection of mycoplasmas. Isr J Med Sci. 1987 Jun;23(6):742–746. [Abstract] [Google Scholar]
  • Giovannoni SJ, DeLong EF, Olsen GJ, Pace NR. Phylogenetic group-specific oligodeoxynucleotide probes for identification of single microbial cells. J Bacteriol. 1988 Feb;170(2):720–726. [Europe PMC free article] [Abstract] [Google Scholar]
  • Wilson KH, Blitchington R, Hindenach B, Greene RC. Species-specific oligonucleotide probes for rRNA of Clostridium difficile and related species. J Clin Microbiol. 1988 Dec;26(12):2484–2488. [Europe PMC free article] [Abstract] [Google Scholar]
  • DeLong EF, Wickham GS, Pace NR. Phylogenetic stains: ribosomal RNA-based probes for the identification of single cells. Science. 1989 Mar 10;243(4896):1360–1363. [Abstract] [Google Scholar]
Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

Full text links 

Read article at publisher's site: https://doi.org/10.1093/nar/17.19.7843

Read article for free, from open access legal sources, via Unpaywall: https://europepmc.org/articles/pmc334891?pdf=renderUnpaywall

Citations & impact 

Impact metrics

1,253
Citations
Jump to Citations
2
Data citations
Jump to Data

Citations of article over time

Alternative metrics

Altmetric item for https://www.altmetric.com/details/41181298
Altmetric
Discover the attention surrounding your research
https://www.altmetric.com/details/41181298

Article citations

Go to all (1,253) article citations

Data 

Data behind the article

This data has been text mined from the article, or deposited into data resources.

BioStudies: supplemental material and supporting data

Similar Articles 

To arrive at the top five similar articles we use a word-weighted algorithm to compare words from the Title and Abstract of each citation.