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Applied and Environmental Microbiology, February 2006, p. 1355-1363, Vol. 72, No. 2
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.2.1355-1363.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Unstable Lysogeny and Pseudolysogeny in Vibrio harveyi Siphovirus-Like Phage 1

Krit Khemayan,^1,2 Tirasak Pasharawipas,³ Orapim Puiprom,^1,2 Siriporn Sriurairatana,¹ Orasa Suthienkul,⁴ and Timothy W. Flegel^1,2*

Centex Shrimp,¹ Department of Biotechnology, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok 10400, Thailand,² Department of Microbiology, Faculty of Science, Rangsit University, Pathumthani 12000, Thailand,³ Department of Microbiology, Faculty of Public Health, Mahidol University, Ratchawithi Road, Bangkok 10400, Thailand⁴

Received 30 August 2005/ Accepted 21 November 2005

Exposure of Vibrio harveyi (strain VH1114) to V. harveyi siphovirus-like phage 1 (VHS1) resulted in the production of a low percentage of lysogenized clones of variable stability. These were retrieved most easily as small colonies within dot plaques. Analysis revealed that VHS1 prophage was most likely carried by VH1114 as an episome rather than integrated into the host chromosome. In the late exponential growth phase, lysogenized VH1114 continuously produced VHS1 but also gave rise to a large number of cured progeny. The absence of phage DNA in the cured progeny was confirmed by the absence of VHS1 DNA in Southern blot and PCR assays. Curiously, these very stable, cured subclones did not show the parental phenotype of clear plaques with VHS1 but instead showed turbid plaques, both in overlaid lawns and in dot plaque assays. This phenotypic difference from the original parental isolate suggested that transient lysogeny by VHS1 had resulted in a stable genetic change in the cured clones. Such clones may be called pseudolysogens (i.e., false lysogens), since they have undergone transient lysogeny and have retained some resistance to full lytic phage development, despite the loss of viable or detectable prophage.

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* Corresponding author. Mailing address: Centex Shrimp and Department of Biotechnology, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok 10400, Thailand. Phone: 6622015870. Fax: 6623547344. E-mail: sctwf{at}mahidol.ac.th.

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

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Applied and Environmental Microbiology, February 2006, p. 1355-1363, Vol. 72, No. 2
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.2.1355-1363.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

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