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Journal of Bacteriology, July 2009, p. 4458-4464, Vol. 191, No. 13
0021-9193/09/$08.00+0     doi:10.1128/JB.00368-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Site-Specific Recombination in the Cyanobacterium Anabaena sp. Strain PCC 7120 Catalyzed by the Integrase of Coliphage HK022{triangledown}

Olga Melnikov,1* Arieh Zaritsky,1 Aliza Zarka,2 Sammy Boussiba,2 Natalia Malchin,3 Ezra Yagil,3 and Mikhail Kolot3

Department of Life Sciences, Ben-Gurion University of the Negev, P.O. Box 653, Be'er-Sheva 84105, Israel,1 Microalgal Biotechnology Laboratory, Blaustein Institute for Desert Research, Ben-Gurion University at Sede-Boker, Sede-Boker 84990, Israel,2 Department of Biochemistry, Tel-Aviv University, Tel-Aviv 69978, Israel3

Received 17 March 2009/ Accepted 27 April 2009

The integrase (Int) of the {lambda}-like coliphage HK022 catalyzes the site-specific integration and excision of the phage DNA into and from the chromosome of its host, Escherichia coli. Int recognizes two different pairs of recombining sites attP x attB and attL x attR for integration and excision, respectively. This system was adapted to the cyanobacterium Anabaena sp. strain PCC 7120 as a potential tool for site-specific gene manipulations in the cyanobacterium. Two plasmids were consecutively cointroduced by conjugation into Anabaena cells, one plasmid that expresses HK022 Int recombinase and the other plasmid that carries the excision substrate PglnA-attL-T1/T2-attR-lacZ, where T1/T2 are the strong transcription terminators of rrnB, to prevent expression of the lacZ reporter under the constitutive promoter PglnA. The Int-catalyzed site-specific recombination reaction was monitored by the expression of lacZ emanating as a result of T1/T2 excision. Int catalyzed the site-specific excision reaction in Anabaena cells when its substrate was located either on the plasmid or on the chromosome with no need to supply an accessory protein, such as integration host factor and excisionase (Xis), which are indispensable for this reaction in its host, E. coli.

* Corresponding author. Mailing address: Department of Life Sciences, Ben-Gurion University of the Negev, P.O. Box 653, Be'er-Sheva 84105, Israel. Phone: 972-8-6461712. Fax: 972-8-6278951. E-mail: burgazla{at}bgu.ac.il

{triangledown} Published ahead of print on 8 May 2009.

Journal of Bacteriology, July 2009, p. 4458-4464, Vol. 191, No. 13
0021-9193/09/$08.00+0     doi:10.1128/JB.00368-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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