Nucleic Acids Research, 1989, Vol. 17, No. 19 7843-7853
© 1989
MOLECULAR BIOLOGY |
Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA
Institut für Medizinische Mikrobiologie, Medizinische Hochschule Hannover Konstanty-Gutschow-Strasse 8, D-3000 Hannover 61 1GBF (Gesellschaft für Biotechnologische Forschung), Mascheroder Weg 1, D-3300 Braunschweig, FRG
*To whom correspondence should be addressed
Received August 2, 1989. Accepted September 4, 1989.
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.
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