林肯链霉菌双亲灭活原生质体融合的研究

Study on Fusion of Biparental Inactivated Protoplasts of Streptomyces lincolnensis

分别以紫外线、热灭活林肯链霉菌 94 7和 95 0 2原生质体 ,然后进行灭活双亲的原生质体融合 ,从 1 6株融合子筛选到林肯霉素高产株。用双亲的互补营养缺陷型对林肯链霉菌原生质体的制备、融合、再生的部分条件进行了研究。发现含 0 .4 %Gly和 34 %蔗糖的SM培养基最适于实验菌株原生质体的制备、再生。聚乙二醇 (PEG)分子量对原生质体融合影响不大 ,其在P缓冲液中的浓度却很重要。含 5 0

Streptomyces lincolnensis 947 and 9502 protoplasts were treated by UV and heat respectively to make them inactivated and then fusion between biparental inactivated protoplasts was carried out. Among 16 fusants, a highly lincomycin producing strain was obtained. Some relative conditions for preparation, fusion and regeneration of the S.lincolnensis protoplasts were studied. It was proved that SM medium containing 0.4% Gly and 34% sucrose was optimal condition for preparation and regeneration of the protoplasts. PEG in different molecular weight tested in the experiment had little influence on fusion of the protoplasts, but its concentration in P buffer was of importance. PEG at a concentration of about 50% in P buffer was of benefit to the protoplast fusion.