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Appl Environ Microbiol, June 1998, p. 2061-2064, Vol. 64, No. 6
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Development of a Genetic Transformation System for an Alga-Lysing Bacterium

Junichi Kato,1,* Junya Amie,1 Yoshinori Murata,1 Akio Kuroda,1 Atsushi Mitsutani,2 and Hisao Ohtake1

Department of Fermentation Technology, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8527,1 and Department of Biology and Aquaculture, National Fisheries University, Shimonoseki, Yamaguchi 756-6595,2 Japan

Received 14 January 1998/Accepted 1 April 1998

Four marine bacteria, Alteromonas sp. strains A27, A28, A29, and A30, that lyse the diatom Skeletonema costatum NIES-324 were isolated from coastal seawater samples. They were also able to lyse the diatoms Thalassiosira sp. and Eucampia zodiacs and the raphidophycean flagellate Chattonella antiqua. Cryptic indigenous plasmids, designated pAS28 and pAS29, were detected in Alteromonas sp. strains A28 and A29, respectively. These plasmids appeared to be similar based on size and restriction site analysis. A shuttle vector that replicates in Escherichia coli and Alteromonas sp. strain A28 was constructed by fusing pAS28 and E. coli vector pCRIIc. The 16-kbp chimeric plasmid, designated pASS1, had the ability to transform strain A28 at a frequency of 106 transformants per µg of DNA. Deletion analysis of pASS1 showed that the 4.7-kb EcoRI-HindIII region of pAS28 was essential for plasmid maintenance in strain A28. This EcoRI-HindIII fragment contained an open reading frame which appeared to encode a 708-amino-acid protein.

* Corresponding author. Mailing address: Department of Fermentation Technology, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8527, Japan. Phone: 81-824-24-7757. Fax: 81-824-22-3758. E-mail: jun{at}ipc.hiroshima-u.ac.jp.

Appl Environ Microbiol, June 1998, p. 2061-2064, Vol. 64, No. 6
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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