The Spheroplast Formation and Regeneration of Pseudomonas spp.

Pseudomonas spp.의 원형질체 생성과 재생

  • 이주실 (고려대학교 이과대학 생물학과) ;
  • 이영록 (고려대학교 이과대학 생물학과)
  • Published : 1986.03.01


The optimal conditions for the formation and the regeneration of Pseudomonas spheroplast were measured. Pseudomonas spp. cells were transformed to spheroplasts from 99.0% to 99.9% by treatment of $100{\mu}g/ml$ lysozyme and 10mM EDTA at room temperature. The optimal pH for the spheroplast formation was pH 8.0. Magnecium chloride, calcium chloride and streptomycin were effective on the stabilization of Pseudomonas spheroplast, while $Mg^+\;and\;Na^+$ ions were effective on the formation of Pseudonomas spheroplast. Rich Regeneration Medium was used for the regeneration of Pseudonomas spheroplast. To improve regeneration frequency, Bovin Serum Albumine and cationic ions were added to the spheroplast dilution beffer and regeneration environment respectively. Treatment of 20mM calcium chloride in ehr Rich Regeneration Medium could improve the yield of regenerants as much as 28-fold. Treatment of 1% Bovin Serum Albumine in the spgeroplast formation and dilution buffer increased the yield of regenerants to 10-fold. Also, the regeneration frequency was improved to 14-fold shen Rich Regeneration Meidum containing 0.5% Gelatin was used for regeneration as well as 1% Bovin Serum Albumine.