Amplification and cloning of the Mycobacterium tuberculosis dnaA gene.
Gene
; 163(1): 75-9, 1995 Sep 22.
Article
em En
| MEDLINE
| ID: mdl-7557482
ABSTRACT
To identify and subsequently clone the gene encoding the DnaA protein, degenerate oligodeoxyribonucleotide (oligo) primers targeted against two highly conserved domains of the eubacterial DnaA were used to amplify a 780-bp DNA region spanning the two primers from genomic DNA preparations of Mycobacterium tuberculosis (Mt), M. bovis (Mb) and M. avium (Ma). Nucleotide (nt) sequences and deduced amino acid (aa) sequences of these fragments revealed homologies with each other and with the corresponding regions from other bacteria. Using an oligo specific to Mt dnaA as a probe, the Mt genomic DNA cosmid libraries propagated in Escherichia coli were screened and a cosmid DNA clone hybridizing with the oligo was identified. Furthermore, a 5-kb DNA fragment containing the Mt dnaA was subcloned into a pUC18 vector.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas de Bactérias
/
Proteínas de Ligação a DNA
/
Genes Bacterianos
/
Mycobacterium tuberculosis
Idioma:
En
Revista:
Gene
Ano de publicação:
1995
Tipo de documento:
Article
País de afiliação:
Estados Unidos