【摘要】 测定了球形红假单胞菌原核细胞与酿酒酵母真核细胞的原生质体形成、再生及融合最佳组合条件。以溶菌酶ＥＤＴＡ反应系统处理球形红假单胞菌Ｐ９４７９，最适脱壁条件为：溶菌酶浓度０５ｍｇ／ｍｌ，ＥＤＴＡ浓度为０２％，蔗糖浓度２０％，酶作用时间为４０ｍｉｎ；此条件下原生质体形率为７８９％，再生率为１１２％。用蜗牛酶巯基乙醇反应系统处理酿酒酵母Ｙ９４０７，最适脱壁条件为：蜗牛酶浓度１０％，巯基乙醇浓度０１％，蔗糖浓度２０％，酶作用时间为３０ｍｉｎ；此条件下酵母原生质体形成率为９９８％，再生率为９７％。用聚乙二醇诱导Ｐ９４７９与Ｙ９４０７的原生质体发生融合，当聚乙二醇的浓度为３０％，Ｃａ２＋浓度为５０ｍｍｏｌ／Ｌ，ｐＨ为６５，时间为１０ｍｉｎ时，有最高的融合率为７６×１０－６。更多还原

【Abstract】 The protoplasts of S.cervisiae and R.sphaeroides were fused in the different conditions.The fusants belong to the hybridization product between eukaryote and prokaryote cells.The level of factors for R.sphaeroides P9479 to form protoplast were lysozyme 0.5mg/ml,EDTA 0.2%,sucrose 20%,30℃,pH 8.0 and 40 min reaction time;and snail enzyme 1.0%,sulghydryl ethanol 0 1%,sucrose 20%,30℃,pH 4.8 and 30 min reaction time for S.cerevisiae to form protoplasts.The fusion between R.sphaeroides and S.cerevisiae protoplasts needed the suitable reaction conditions which were PEG(MW=6000) 30%,Ca 2+ 50mmol/L,30℃ and 10min reaction time.The highest level of protoplast formation and regeneration rates could be obtained which were 78.9% and 11.2% for R.sphaeroides ,99.8% and 9.7% for S.cerevisiae ,and the highest fusion frequency between R.sphaeroides and S.cerevisiae protoplast was 7.6×10 -6 .更多还原