摘要
研究了热带假丝酵母野生株１０４５及其突变株ＳＤ１的原生质体分离、电击诱导融合的诸条件以及融合子的再生。得出了原生质体的最佳生成条件。用９９９ｋＨｚ，２４０～４００ Ｖ／ｃｍ的高频交变电场使原生质体横向电泳形成珠串，以脉宽 ４０ μｓ，幅值 １６００～２ ８００ Ｖ／ｃｍ的直流脉冲可使原生质体融合，净融合率达６５％。采用较低幅值的多个脉冲有利于融合。电击液中加入ＣａＡｃ场、ＭｇＡｃ２及白蛋白可大大提高融合率，用光刻镀金方法设计了适用于微生物的多电极融合小室。对膜融合机制进行了探讨。
The isolation, electrofusion and regeneration of protoplasts from Candida tropicalis wild strain 1045 and its mutant SD1 were investigated. By using early exponential phase cells of 5 x 107 concentration under the action of 2% snail enzyme and 0.1 % β-mercaptoethanol, the rate of protoplast formation was observed to be about 90 %, while the rate of regeneration about 79%. The fusion of protoplasts was induced by the A. C. field of 999kHz, 240-400 V/cm and the D.C. pulses of 40μs, 1600-2 600 V/cm. Under optimum conditions, the fusion yield was 65%. Several factors which might influence the fusion yield were tested and discussed. The fusion chamber was improved to obtain higher fusion yield. The experimental results show that the fusion yield can be raised by using more pulses of lower voltage and it can also be raised greatly by adding 0. 1 mmol/L CaAc2, 0.5 mmol/L MgAc2 and 1 g/L albumin.
出处
《清华大学学报（自然科学版）》
EI
CAS
CSCD
北大核心
1990年第6期60-67,共8页
Journal of Tsinghua University(Science and Technology)
关键词
原生质体
酵母菌
电场诱导
融合
protoplast, yeast, electric field-induced fusion
