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Journal of Bacteriology, January 2000, p. 30-37, Vol. 182, No. 1
0021-9193/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Identification and Characterization of an Active Plasmid Partition Mechanism for the Novel Lactococcus lactis Plasmid pCI2000

Karen Kearney,1,2,3 Gerald F. Fitzgerald,1,2,3,* and Jos F. M. L. Seegers3

Department of Microbiology1 and Food Science and Technology2 and National Food Biotechnology Centre,3 University College, National University of Ireland, Cork, Ireland

Received 18 June 1999/Accepted 7 October 1999

The replication region of the lactococcal plasmid pCI2000 was subcloned and analyzed. The nucleotide sequence of one 5.6-kb EcoRI fragment which was capable of supporting replication when cloned on a replication probe vector revealed the presence of seven putative open reading frames (ORFs). One ORF exhibited significant homology to several replication proteins from plasmids considered to replicate via a theta mode. Deletion analysis showed that this ORF, designated repA, is indeed required for replication. The results also suggest that the origin of replication is located outside repA. Upstream and divergently transcribed from repA, an ORF that showed significant (48 to 64%) homology to a number of proteins that are required for faithful segregation of chromosomal or plasmid DNA of gram-negative bacteria was identified. Gene interruption and transcomplementation experiments showed that this ORF, designated parA, is required for stable inheritance of pCI2000 and is active in trans. This is the first example of such a partitioning mechanism for plasmids in gram-positive bacteria.

* Corresponding author. Mailing address: Department of Microbiology, University College, National University of Ireland, Western Rd., Cork, Ireland. Phone: 353 21 902730. Fax: 353 21 903101. E-mail: g.fitzgerald{at}ucc.ie.

Journal of Bacteriology, January 2000, p. 30-37, Vol. 182, No. 1
0021-9193/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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