SAP2 mRNA表达在白色假丝酵母菌二相性中的研究

Expression of SAP2 mRNA in dimorphism of Candida albicans

目的通过对编码白色假丝酵母菌分泌型天冬氨酸蛋白酶2(SAP2)基因在该菌酵母相和菌丝相中的mRNA表达水平的比较,从分子生物学角度探讨编码白色假丝酵母菌侵袭性酶的重要毒力基因在白色假丝酵母菌二相性中的表达差异。方法采用RNAiso Plus试剂联合玻璃珠破壁方法,分别提取白色假丝酵母菌的酵母相和菌丝相的总RNA;应用半定量RT-PCR技术,比较SAP2基因在白色假丝酵母菌酵母相和菌丝相中mRNA水平的表达差异。结果提取的白色假丝酵母菌酵母相及菌丝相总RNA电泳结果均可见较为清晰的28S、18S、5S3个条带;运用半定量RT-PCR技术,得出SAP2基因在酵母相和菌丝相内mRNA的相对表达量分别为0.789±0.039,0.927±0.062,两者比较差异有统计学意义(P<0.05)。结论采用RNAiso Plus试剂联合玻璃珠破壁方法提取的白色假丝酵母菌酵母相和菌丝相总RNA的浓度和纯度较高、完整性好;SAP2基因在白色假丝酵母菌菌丝相mRNA表达水平明显高于酵母相。

OBJECTIVE To study the mRNA expression of SAP2 gene encoding the secreted aspartyl proteinase 2 in the yeast form and hyphal form of Candida albicans and discuss the expression of virulence gene encoding invasive enzymes in dimorphism of C. albicans by the perspective of molecular biology. METHODS The total RNA of yeast form and hyphal form of C. albicans were extracted by RNAiso Plus reagent combined ~vith glass beads method~ the differences in the mRNA expression of SAP2 were observed by semi-quantitative RT-PCR. RESULTS The result of electrophoresis of the extracted total RNA in yeast form and hyphal form showed that three clear bands appeared： 28s rRNA, 18s Rrna, and 5s rRNA, indicating that the extracted total RNA in yeast form and hyphal form were both integrated. The sem-quantitative RT-PCT showed that the relative expression of mRNA in SAP2 gene was （0. 789 ~ 0. 039） in yeast form, （0. 927 ~ 0. 062） in hyphal form, the difference was significant （P〈0.05）. CONCLUSION The RNAiso Plus reagent combined with glass beads can extract the total RNA of high purity , concentration, and integrity~ the level of expression of SAP2 mRNA is significantly higher in the hyphal form than in the yeast {orm o{ C. albicans.