A Rapid Procedure for Screening and Isolation of Various Sizes of Plasmid DNA in Serovars of Bacillus thuringiensis

Bacillus turingiensis 변종(變種)들로부터의 Plasmid DNA 추출(抽出) 및 분리(分離)

  • LEE, YUNG KEUN (Sericultural Experiment Station, Office of Rural Development) ;
  • Faust, Robert M. (Insect Pathology Laboratory, ARS, USDA) ;
  • KANG, SEOK KWON (Department of Sericulture, College of Agriculture, Seoul National University) ;
  • McCawley, Patricia E. (Insect Pathology Laboratory, ARS, USDA) ;
  • Meyers-Dowling, Carol L. (Insect Pathology Laboratory, ARS, USDA)
  • 이영근 (농촌진흥청, 잠업시험장) ;
  • 로베트 엠 파우스트 (미농무성, 농업연구소, 곤충병리학연구실) ;
  • 강석권 (서울대학교 농과대학 잠사학과) ;
  • 페트리시아 이 멕콜리 (미농무성, 농업연구소, 곤충병리학연구실) ;
  • 케롤 엘 메이어-다운링 (미농무성, 농업연구소, 곤충병리학연구실)
  • Published : 1985.04.30

Abstract

The use of a modified procedure for the isolation of extrachromosomal DNA of low to high molecular weight, followed by agarose gel electrophoresis of the crude lysates, provided a simple screening procedure for detecting plasmids ranging in molecular weights from approximately 1 to more than 135 megadaltons from serovars of Bacillus thuringiensis. The procedure provides for a relatively large-volume stable lysate for isolation of plasmids for restriction endonuclease mapping and cloning procedures. The method was used for screening of plasm ids in 6 differenentially effective serovars of B. thuringiensis toxic to dipteran and lepidopteran insects. Relatively large plasmid DNAs of masses above 50 megadaltons (Mdal) were isolated from all of the serovars examined using this technique. The number of extrachromosomal DNAs detected in serovars of B. thuringiensis was 8 for israelensis, 10 for kurstaki, 13 for aizawai, 2 for dendrolimus, 1 for finitimus, and 6 for yunnanensis. Smaller plasmid DNAs were isolated in four of the six serovars that ranged in mass down to approximately 2 Mdal.

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