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Cloning and sequencing of the replication origin ( ori C) of the Spiroplasma citri chromosome and construction of autonomously replicating artificial plasmids

Identifieur interne : 004470 ( Main/Exploration ); précédent : 004469; suivant : 004471

Cloning and sequencing of the replication origin ( ori C) of the Spiroplasma citri chromosome and construction of autonomously replicating artificial plasmids

Auteurs : Fengchun Ye [France] ; Joël Renaudin [France] ; Joseph-Marie Bové [France] ; Frédéric Laigret [France]

Source :

RBID : ISTEX:F1074516372ED7167FB9C23431CFD906E0643050

English descriptors

Abstract

Abstract: A 5.6-kbp fragment ofSpiroplasma citri DNA containing thednaA gene has been cloned and sequenced. Nucleotide sequence analysis shows that this fragment harbors the genes for the replication initiator protein (dnaA), the beta subunit of DNA polymerase III (dnaN), and the DNA gyrase subunits A and B (gyrA andgyrB). The arrangement of these genes,dnaA-dnaN-gyrB-gyrA, is similar to that found in all Gram-positive bacterial genomes studied so far, except that norecF gene was found betweendnaN andgyrB. Several DnaA-box consensus sequences were found upstream ofdnaA and in thednaA-dnaN intergenic region. ThednaA region with the flanking DnaA-boxes and the tetracycline resistance determinant,tetM, were linked into a circular recombinant DNA. This DNA was able to replicate autonomously when introduced by electroporation intoS. citri cells. These experiments show that thednaA region with the DnaA-boxes is the origin of replication ofS. citri and can be used to construct gene vectors.

Url:
DOI: 10.1007/BF01570187


Affiliations:


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Le document en format XML

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<term>Bacillus subtilis</term>
<term>Bacterial chromosome</term>
<term>Buchnera aphidicola</term>
<term>Capricolum</term>
<term>Chromosome</term>
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<term>Coli</term>
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<term>Pneumoniae</term>
<term>Pot1</term>
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<term>Primer</term>
<term>Recf gene</term>
<term>Replication</term>
<term>Replication origin</term>
<term>Sequencing</term>
<term>Spiroplasma</term>
<term>Spiroplasma citri</term>
<term>Spiroplasma citri genome</term>
<term>Subtilis</term>
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<term>Taqi fragment</term>
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<div type="abstract" xml:lang="en">Abstract: A 5.6-kbp fragment ofSpiroplasma citri DNA containing thednaA gene has been cloned and sequenced. Nucleotide sequence analysis shows that this fragment harbors the genes for the replication initiator protein (dnaA), the beta subunit of DNA polymerase III (dnaN), and the DNA gyrase subunits A and B (gyrA andgyrB). The arrangement of these genes,dnaA-dnaN-gyrB-gyrA, is similar to that found in all Gram-positive bacterial genomes studied so far, except that norecF gene was found betweendnaN andgyrB. Several DnaA-box consensus sequences were found upstream ofdnaA and in thednaA-dnaN intergenic region. ThednaA region with the flanking DnaA-boxes and the tetracycline resistance determinant,tetM, were linked into a circular recombinant DNA. This DNA was able to replicate autonomously when introduced by electroporation intoS. citri cells. These experiments show that thednaA region with the DnaA-boxes is the origin of replication ofS. citri and can be used to construct gene vectors.</div>
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