食品工业科技2012，Vol.33Issue(12)：233-238,6.
同种接合型酵母菌株Y2HGold的原生质体融合研究
Protoplast fusion between the same mating type strains of Y2HGold
摘要
Abstract
In order to avoid the limitations of sexual fusion and find out the potential value of asexual fusion between yeast strains of same mating type,the MatchmakerTM Gold Yeast Two-Hybrid System strain Y2HGold were chosen as the material,using the plasmid,the leucine-deficient and tryptophan-deficient strains of Y2HGold were successfully constructed;using orthogonal test with four factors and three levels by enzymolysis method,the preparation protocol of deficient-bacteria protoplasts was concluded as：enzymolysis temperature of 30℃,80min processing time,pre-treatment agent containing 0.1%（w/v）mercaptoethanol and 0.1%（w/v）EDTA-2Na,hydrolyzate a mixture of 1.5%（w/v）and 1.5% snail enzyme（w/v）cellulase,rendering the protoplast preparation and regeneration rate 82% and 37% respectively.Finally,using PEG-induced fusion method and the same orthogonal test,we obtained the requirements about asexual fusion of Y2HGold：35% PEG concentration,temperature as 25℃ and 40min and pH5.8.Sub-clones we selected were genetically stable.关键词
酵母/原生质体/无性融合/无性融合菌株Key words
yeast/protoplast/asexual fusion/asexual fusion strains分类
轻工纺织引用本文复制引用
常登龙,洪玉,杨永军,张贺翠,薛丽琰,杨昆,朱利泉,王小佳..同种接合型酵母菌株Y2HGold的原生质体融合研究[J].食品工业科技,2012,33(12):233-238,6.基金项目
重庆市自然基金重点项目 （）
国家自然科学基金项目 （）
国家大学生创新计划项目 （）
