Rastogi N, David H L
J Gen Microbiol. 1981 May;124(1):71-9. doi: 10.1099/00221287-124-1-71.
Wall-deficient forms of Mycobacterium aurum were prepared by agitating the cells during exponential growth with D-cycloserine, glycine, lysozyme, EDTA and LiCl for approximately the time of three cell divisions (18 h). Wall-deficient forms were then converted to spheroplasts by gentle stirring with lysozyme and EDTA in a Tris/HCl buffer containing sucrose until all the cells appeared spherical by phase contrast microscopy. Subsequent lysis by nucleases followed by osmotic shock produced membrane vesicles. Ultrastructural and chemical properties of the spheroplasts and membrane vesicles are described. The spheroplasts were susceptible to lysis by 0.25% (w/v) sodium dodecyl sulphate and were permeable to certain enzyme substrates.
通过在指数生长期用D-环丝氨酸、甘氨酸、溶菌酶、乙二胺四乙酸（EDTA）和氯化锂搅拌细胞约三个细胞分裂时间（18小时），制备出金色分枝杆菌的细胞壁缺陷型。然后，在含有蔗糖的Tris/HCl缓冲液中用溶菌酶和EDTA轻轻搅拌，将细胞壁缺陷型转化为原生质球，直到相差显微镜下所有细胞都呈球形。随后用核酸酶裂解，接着进行渗透休克，产生膜泡。描述了原生质球和膜泡的超微结构及化学性质。原生质球易被0.25%（w/v）十二烷基硫酸钠裂解，并且对某些酶底物具有通透性。
