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J. Microbiol. Biotechnol. 2000; 10(3): 405-409

Published online June 28, 2000

Copyright © The Korean Society for Microbiology and Biotechnology.

Analysis of the Genome of Symbiobacterium toebii by Pulsed - Field Gel Electrophoresis

Seung-Pyo Hong *, Jong-Hoon Park 1, Yong Sung Kim 1, Hae-Jun Hwang 1, Sung-Keun Rhee 1, Seung-Goo Lee 1, Moon-Hee Sung 1 and Nobuyoshi Esak 2

Environmental Bioresources Laboratory Korea Research Institute of Bioscience and Biotechnology (KRIBB) Yusong Taejon Korea, 1Genome Research Center Korea Research Institute of Bioscience and Biotechnology (KRIBB) Yusong Taejon Korea, 2Molecular Microbial Science ICR Kyoto University Uji Kyoto 611-0011 Japan

Abstract

We have studied the genome of an obligately commensal thermophile, Symbiobacterium toebii. The chromosome was extracted from pure cultures of S. toebii recently established. Total DNA of S. toebii was resolved by pulsed-field gel electrophoresis (PFGE) into discrete numbers of fragments by digenstion with the endonuclease SspI, SpeI, XbaI, and HpaI. Estimated sizes of fragments produced by the four enzymes and their sum consistently yielded a total genome size of 2.8 Mb. Because restriction endonucleases NotI and SwaI, recognizing 8 bp, released too many fragments, these enzymes could not be used for the estimation of the genome size. Considering no mobility of undigested genome under PFGE, the genome of S. toebii appears to be circular. The presence of extrachromosomal DNA in S. toebii was excluded by the results of the conventional 1% agarose gel electrophoresis and the field inversion gel electrophoresis of undigested S. toebii DNA.

Keywords: Symbiobacteriumtoebii, obligatelycommensalthermophile, genome