阿维链霉菌原生质体制备方法的研究

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。

Preparation Methods for Protoplast during Breeding of Streptomyces avermitilis

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/min 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.