미생물학회지 (Korean Journal of Microbiology)

  • 제30권4호
  • /
  • Pages.286-290
  • /
  • 1992
  • /
  • 0440-2413(pISSN)
  • /
  • 2383-9902(eISSN)
한국미생물학회 (The Microbiological Society of Korea)
권호 표지

Siderophore를 생성하는 Fluorescent Pseudomonads의 분리, 동정 및 돌연번이 유기

Identification of Fluorescent Pseudomonads Producing Siderophore and Construction of Siderophore Biosynthesis Defective Mutant

  • Park, Yeal (Department of Genetic Engineering) ;
  • Kim, Hyun Hee (Department of Genetic Science, Graduate School, Chosun University) ;
  • Myeong-gu Yeo (Department of Genetic Science, Graduate School, Chosun University) ;
  • Young-woo Seo (Department of Genetic Science, Graduate School, Chosun University) ;
  • Han-cheol Koh (Department of Genetic Science, Graduate School, Chosun University) ;
  • Young-gi Yang (Department of Genetic Engineering) ;
  • Hyeon-Sook Cheong (Department of Genetic Engineering) ;
  • Sung-jun Kim (Department of Genetic Engineering)
  • 발행 : 1992.08.01
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초록

광주근교 지역의 근권 토양으로부터 cetrimide agar medium을 이용하여 형광성 pseudomonads를 분리하였고, CAS medium에서 siderophore의 생성능력이 우수한 pseudomonads만을 분리하여 생리화학적인 실험을 수행하였다. Kanamycin-sensitive pseudomonads를 Tn5를 이용한 mutagenesis를 실시하여 Kanamycin에 내성을 갖는 transconjugants를 선별하였고, siderophore 생합성을 하지 못하는 돌연변이주를 선별하기 위하여 CAS medium에서 yellow hallow를 형성하지 못하거나 King's B medium에서 형광성을 나타내지 못하는 colony를 선별하였다. 선별된 mutants들의 genomic DNA에 Tn5가 삽입되었는지를 확인하기 위하여 Southern blot hybridization을 실시한 결과 intact Tn5에 homology를 나타내는 하나의 single band를 확인하였다.

The present study was performed to isolate the fluorescent pseudomonads from Kwang-Ju soil and to construct a mutant strain defective in siderophore biosynthesis. The siderophore-secreting pseudomonads were screened on Blue agar (Chrome Azuol S agar) plates and one strain of them was designated to Pseudominas fluorescens (P. fluorescens) PY002. To construct a mutant defective in siderophore biosynthesis, P. fluorescens PY002 was randomly mutagenized with a transposon Tn5. The location of Tn5 integrated into chromosomal of the mutants strain was determined by Southern blot analysis. The mutagenized strain showed non-fluorescent on a King's B agar plate and were defective in iron (III) acquisition ability.

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