Isolation and Direct Complete Nucleotide Determination of Entire Genes. Characterization of a Gene Coding for 16S Ribosomal RNA

Nucleic Acids Res. 1989 Oct 11;17(19):7843-53. doi: 10.1093/nar/17.19.7843.

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.

Publication types

  • Comparative Study

MeSH terms

  • Bacteria / genetics*
  • Base Sequence
  • DNA-Directed DNA Polymerase
  • Genes, Bacterial*
  • Molecular Sequence Data
  • Mycobacterium / genetics*
  • Polymerase Chain Reaction
  • RNA, Ribosomal / genetics*
  • RNA, Ribosomal, 16S / genetics*
  • Sequence Homology, Nucleic Acid

Substances

  • RNA, Ribosomal
  • RNA, Ribosomal, 16S
  • DNA-Directed DNA Polymerase

Associated data

  • GENBANK/X15916