摘要
对阿维链霉菌原生质体制备进行了研究,结果表明:阿维链霉菌原生质体制备的最佳条件为在35 mL含0.5%甘氨酸的YEME培养基中接种孢子悬液,28℃、200 r/min培养40 h;3 000 r/min收集菌丝体,用10.3%蔗糖溶液清洗2次;溶菌酶浓度为4mg/mL的P缓冲液悬浮菌丝体,30℃下处理50 min完成原生质体制备。以50%PEG1000为促融剂,原生质体融合率最高。
This research aimed to study the preparation for protoplast of Streptomyces avermitilis. The results showed that the optimum conditions for preparation of protoplast are as follows: at first, inoculation of spore suspending liquid to YEME culture medium which containing 35 mL of 0.5% glycin, and then cultured at 28℃ with 200 r/rain for 40 h; then, collecting mycelia with 3 000 r/min, and then the collected mycelia were washed by 10.3% sucrose solution for two times; finally, the cleaned mycelia were suspended in P buffer with 4 mg/mL of lysozyme at 30℃ for 50 min to finish the preparation of protoplast. When 50% PEG 1 000 was used as fusion promoting agent, the fusion rate of protoplasts was the highest.
出处
《湖南农业科学》
2012年第4期14-16,共3页
Hunan Agricultural Sciences
关键词
阿维链霉菌
阿维菌素
原生质体制备
Streptomyces avermitilis
avermectins
protoplast preparation