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PMID: 7764984 Published · ppublish English Journal Article

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

Current microbiology ·Vol. 29 ·No. 1 ·1994-07-00 ·Pages 23-9

Ye F, Renaudin J, Bové JM, Laigret F

Abstract

A 5.6-kbp fragment of Spiroplasma citri DNA containing the dnaA 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 and gyrB). 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 no recF gene was found between dnaN and gyrB. Several DnaA-box consensus sequences were found upstream of dnaA and in the dnaA-dnaN intergenic region. The dnaA 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 into S. citri cells. These experiments show that the dnaA region with the DnaA-boxes is the origin of replication of S. citri and can be used to construct gene vectors.

Related Genes
dnaA dnaN gyrA gyrB
MeSH Terms
Amino Acid Sequence Bacterial Proteins/genetics Base Sequence Chromosome Mapping Chromosomes, Bacterial Cloning, Molecular Consensus Sequence/genetics DNA Gyrase DNA Polymerase III/genetics DNA Replication/genetics DNA Topoisomerases, Type II/genetics DNA, Bacterial/analysis DNA-Binding Proteins/genetics Genes, Bacterial/genetics Molecular Sequence Data Plasmids/genetics Sequence Analysis, DNA Sequence Homology, Amino Acid Spiroplasma/genetics
Chemicals
Bacterial Proteins DNA, Bacterial DNA-Binding Proteins DnaA protein, Bacteria DNA Polymerase III DNA Gyrase DNA Topoisomerases, Type II
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Ye F
Laboratory of Cellular and Molecular Biology, National Institute of Agronomy Research, Villenave d'Ornon, France.
Renaudin J
Bové J M
Laigret F
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Article Info
Journal
Current microbiology
Abbr.
Curr Microbiol
ISSN
0343-8651
DOI
10.1007/BF01570187
Published
1994-07-00
Pages
23-9
Language
English
Region
United States
NLM ID
7808448
Databases
GENBANK
Z19108
External Links
PubMed source DOI: 10.1007/BF01570187 View journal details
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