Luchkina L A, Bekker M L
Prikl Biokhim Mikrobiol. 1975 Mar-Apr;11(2):264-8.
A method for producing spheroplasts from yeast cells has been developed. The method involves: 1) prefreezing and thawing of cells in the minimal nutrient medium; 2) treatment with the mixture of 86 mM 2-mercaptoethanol, 5% sodium dodecylsulphate and 6% Triton X-100; 3) enzymic digestion by the digestive juice of grape helix. The formation of spheroplasts was controlled by microscopy and measurement of the optic density of the spheroplast suspension in the incubation medium and in 1% sodium dodecylsulphate. The method has been successfully tested on some bakery yeast: Saccharomyces cerevisiae, Saccharomyces paradoxus, Schizosaccharomyces pombe, Saccharomyces globosus, Saccharomyces carlsbergensis.
已开发出一种从酵母细胞制备原生质球的方法。该方法包括:1)在基本营养培养基中对细胞进行预冷冻和解冻;2)用86 mM 2-巯基乙醇、5%十二烷基硫酸钠和6% Triton X-100的混合物处理;3)用葡萄螺旋消化液进行酶解。通过显微镜观察以及测定在孵育培养基和1%十二烷基硫酸钠中原生质球悬浮液的光密度来控制原生质球的形成。该方法已在一些面包酵母上成功测试:酿酒酵母、奇异酵母、粟酒裂殖酵母、球形酵母、卡尔斯伯酵母。