白色念珠菌ERG11突变基因在毕氏酵母中的表达

Expression of Candida albicans point mutant ERG11 gene in Pichia pastoris

目的将含有点突变的白色念珠菌编码基因ERG11克隆至毕氏酵母中进行表达，判断其在白色念珠菌对氟康唑敏感性中的作用。方法将含有点突变的白色念珠菌编码基因ERG11与pPm3．5K载体重组，构建pPiC3．5K／Ca1．6k重组体，经Top10F’鉴定和筛选，电转入毕氏酵母GS115中，经过诱导表达出现目的蛋白后，进行药物敏感性试验，确定点突变与耐药的关系。结果含Y132H突变转化菌株和含Q474K突变的转化菌株对氟康唑的MIC分别上升16倍和8倍，证实其突变与耐药有关。结论氟康唑耐药可发生于白色念珠菌编码基因ERG11的Y132H与Q474K点突变中。

Objective To study the relation between the point mutation of Candida albicans ERGll gene and fluconazole susceptibihty. Method ERGll gene was cloned into pPIC3.5K vector, formed the pPIC3.5K/Ca1.6k, then transformed into Pichia pastoris GSll5 and expressed in various condition. Results The susceptibihty test manifested to Y132H mutation increased by 16 folds MIC and Q474K mutation 8 folds respectively. Condusions Resistance to fluconazole may occur through the Y132H point mutation and Q474K point mutation of Candida albicans gene ERGll respectively.