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Applied and Environmental Microbiology, December 2007, p. 7858-7866, Vol. 73, No. 24
0099-2240/07/$08.00+0     doi:10.1128/AEM.01630-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Molecular Dissection of a Bifidobacterial Replicon{triangledown} ,{dagger}

Michelle Cronin, Moritz Knobel, Mary O'Connell-Motherway, Gerald F. Fitzgerald, and Douwe van Sinderen*

Alimentary Pharmabiotic Centre and Department of Microbiology, University College Cork, Western Road, Cork, Ireland

Received 17 July 2007/ Accepted 30 September 2007

The 2.1-kb cryptic plasmid pCIBAO89 from Bifidobacterium asteroides harbors a 1.4-kb segment which is sufficient for its autonomous replication. The segment is divided into two parts, the presumed replication origin, ori89, and the rep gene encoding the putative 41-kDa Rep89 replication initiation protein. This minimal replication region of pCIBAO89 was functionally dissected by transcriptional analyses as well as by DNA-binding studies, and the information obtained was exploited to create a number of Escherichia coli-Bifidobacterium shuttle vectors capable of transforming various bifidobacteria with an efficiency of up to 106 transformants/µg DNA.


* Corresponding author. Mailing address: Department of Microbiology, National University of Ireland, Western Road, Cork, Ireland. Phone: 00 353 214091365. Fax: 00 353 214903101. E-mail: d.vansinderen{at}ucc.ie

{triangledown} Published ahead of print on 26 October 2007.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.


Applied and Environmental Microbiology, December 2007, p. 7858-7866, Vol. 73, No. 24
0099-2240/07/$08.00+0     doi:10.1128/AEM.01630-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.




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