Methodology of Genetic Engineering

遺傳工學의 方法論

  • 이평우 (고려대학교 이공대학 생물학과) ;
  • 윤경하 (고려대학교 이공대학 생물학과) ;
  • 이세영 (연세대학교 분자생물학 연구실)
  • Published : 1975.06.01


The neurotoxin of Clostridium botulinum type B was purified from a liquid culture. The purification steps consist of ammonium sulfate precipitation of whole culture, treatment of Polymin P(0.15%, v/v), gel filtration on Sephadex G-100 at pH5.6 and DEAE-Sephadex charomatography at pH8.0. The procedure recovered 17% of the toxin assayed in the starting culture. The toxin was homogeneous by sodium dodecyl sulfate(SDS)-polyacrylamide gel electrophoresis and had a molecular weight of 163,000. Subunits of 106,000 and 56,000 molecular weight were found when purified toxin was treated with a disulfide-reducing agent and electro phoresed on SDS-polyacrylamide gels.