Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA.
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.
Edwards
U
U
Institut für Medizinische Mikrobiologie, Medizinische Hochschule Hannover, FRG.
Rogall
T
T
Blöcker
H
H
Emde
M
M
Böttger
E C
EC
D001419:Bacteria / Q000235:genetics*; D001483:Base Sequence; D004259:DNA-Directed DNA Polymerase; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012335:RNA, Ribosomal / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid
D012335:RNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D004259:DNA-Directed DNA Polymerase