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

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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.

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Author notes

  1. Fengchun Ye

    Present address: Department of Plant Pathology, Oklahoma State University, 110 Noble Research Center, 74078, Stillwater, OK, USA

Authors and Affiliations

  1. Laboratory of Cellular and Molecular Biology, National Institute of Agronomy Research, and University of Bordeaux II, 33883, Villenave d' Ornon, France

    Fengchun Ye, Joël Renaudin,  Joseph-Marie Bové & Frédéric Laigret

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  1. Fengchun Ye
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  2. Joël Renaudin
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  3. Joseph-Marie Bové
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  4. Frédéric Laigret
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Ye, F., Renaudin, J., Bové, JM. et al. Cloning and sequencing of the replication origin (oriC) of theSpiroplasma citri chromosome and construction of autonomously replicating artificial plasmids. Current Microbiology 29, 23–29 (1994). https://doi.org/10.1007/BF01570187

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