枯草杆菌(B.subtilis)原生质体融合的方法学研究

METHODOLOGY RESEARCH FOR PROTOPLAST FUSION OF Bacillus subtili

利用改良的HM制备液,在适宜的条件下酶解制备亲本株的原生质体,在DNase存在的条件下,用PEG(MW6000)做为聚合剂,经低温短时间处理,将枯草杆菌(B.subtilis)Ki—2—132衍生株与BR_(151)衍生株的原生质体融合。再在适宜的条件下使用改良的DM_3再生培养基,补加适量经处理的人血清白蛋白,同时加入适量适合于枯草杆菌细胞壁再生的引物进行再生,使再生率达到了55～58.15％左右。融合率高达1.82×10^(-2)。筛选出51种不同类型的融合子923株,经传代8代测得其融合子的稳定率达32.46％,从而在工业菌株的选育中使获得高产菌株成为可能。

The parent protoplasts were formed in the improved HM under suitable condition. Treated with PEG(MW6000) under low temperature in short time the protoplasts of derivative B.subtilis Ki-2-132 and those of HR151 were fused in the HMD. With the medium DM3 plus approapriate and treated humam serum albumin and regeneration primers for B.subtilis cell wall, the regeneration rates were raised up to about 55-58.15% and the fusion rate 1.82×10-2, 51 types of fusants were selected from 923 strains, Examing among 8 generates showed the stable rate of fusant was 32.46%. It indicated that aeguiring high yield fusion strains in the industrial microbe breeding was possible.