Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Isolation of Dispersed Mutants from Wild Myxobacteria.

분산 돌연변이 점액세균의 분리

  • 이봉수 (호서대학교 생명과학과) ;
  • 이차율 (호서대학교 생명과학과) ;
  • 조경연 (호서대학교 생명과학과)
  • Published : 2003.12.01
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Abstract

Clumping of cells is one of the major obstacles to culture wild myxobacterial strains in liquid media. In an effort to solve this problem, we tried a method isolating spontaneous mutants that grow dispersed in liquid media from a wild myxobacterial strain. Myxococcus stipitatus KYC1001, a newly isolated strain from Gyearyongsan National Park in Korea, clumps and sticks to the surface of culture vessels as other wild myxobacteria behave in liquid media. Taking an advantage of the characteristics that dispersed mutant cells would grow dispersed while most other wild type cells would clump and stick to the surface of culture vessels, spontaneous dispersed mutants were enriched by repeated subculturing of culture supernatant. A resultant mutant, KYC2001, did not form any clumps nor stick to the surface of culture flasks, but grew completely dispersed in liquid. Meanwhile, three other spontaneous mutants, KYC2002, KYC2003, and KYC2004, shelved partially dispersed phenotype. A major portion of the cells grew dispersed in liquid but they still formed some clumps.

세포들간의 응집 및 배양용기 표면에의 부착은 야생 점액 세균의 액체배양에 큰 장애물로 작용한다. 이러한 장애를 해결하기 위한 한 방안으로 야생 점액세균으로부터 자연적 분산 돌연변이 균주를 얻는 방법을 시도해 보았다. 실험 대상으로 사용한 균주 M. stipitatus KYC1001은 충남 계룡산의 토양에서 분리된 점액세균으로, 다른 점액세균들과 마찬가지로 액체배지 내에서 대부분의 세포들이 응집체를 형성하거나, 배양용기의 표면에 달라붙는다. 본 연구에서는 야생형 균주들이 세포응집체를 형성하거나 배양용기의 표면에 달라붙게 되는 반면 분산 돌연변이 세포들은 배지 내에 자유로이 떠돌아다니게 될 것이라는 특성을 이용하여, 배양액의 상등액만을 30일에 걸쳐 10회 계대 배양함으로서 분산되어 상등액에 존재할 자연적 분산 돌연변이 세포들을 농화 배양하였다. 그 결과, 액체배지에서 완전히 분산되어 성장하는 한 균주, KYC2001과 부분적으로 분산하여 성장하는 세 균주, KYC2002, 2003, 2004를 얻을 수 있었다.

Keywords

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