PLB1基因在白假丝酵母二相性中的表达  

Expression of PLB1gene in dimorphism of Candida albicans

作　　者：罗红梅[1] 苑天红 杨怀[3] LUO Hong-mei;YUAN Tian-hong;YANG Huai(Guiyang Maternal and Child Health Care Hospital,Guiyang,Guizhou 550003,China;不详)

机构地区：[1]贵阳市妇幼保健院新生儿科,贵州贵阳550003 [2]贵州医科大学基础医学院微生物学教研室,贵州贵阳550004 [3]贵州省人民医院医院感染管理科,贵州贵阳550002

出　　处：《中华医院感染学杂志》2023年第16期2427-2431,共5页Chinese Journal of Nosocomiology

基　　金：贵州省科技厅基金资助项目(黔科合J字[2008]2273号)。

摘　　要：目的 通过对编码白假丝酵母磷脂酶B1(PLB1)基因在该菌酵母相和菌丝相中的mRNA和DNA表达水平的比较,从分子生物学角度探讨编码白假丝酵母侵袭性酶的重要毒力基因在该菌二相性中的表达差异,为进一步研究白假丝酵母的遗传变异、致病性与防治提供初步实验依据。方法 采用RNAiso Plus试剂联合玻璃珠破壁方法,分别提取白假丝酵母酵母相和菌丝相的总RNA;应用半定量逆转录聚合酶链式反应(RT-PCR)技术,比较PLB1基因在白假丝酵母酵母相和菌丝相中mRNA水平的表达差异;分别提取白假丝酵母酵母相与菌丝相的总DNA,应用聚合酶链反应-单链构象多态性分析(PCR-SSCP)技术,比较PLB1基因在白假丝酵母酵母相与菌丝相中DNA水平的表达差异。结果 提取的白假丝酵母酵母相及菌丝相总RNA电泳结果均可见较为清晰的28S、18S、5S三条带;运用半定量RT-PCR技术,得出PLB1基因在酵母相和菌丝相内mRNA的相对表达量分别为(0.90±0.04,1.00±0.01),两者比较有统计学差异(P<0.05);提取的白假丝酵母酵母相和菌丝相总DNA,应用PCR-SSCP技术,得到的SSCP图谱显示:PLB1基因在二相中的电泳带型无统计学差异。结论 采用RNAiso Plus试剂联合玻璃珠破壁方法提取的白假丝酵母酵母相和菌丝相总RNA的浓度和纯度较高、完整性好;PLB1基因在白假丝酵母菌丝相mRNA表达水平明显高于酵母相;应用PCR-SSCP技术得到的SSCP图谱显示,所扩增的PLB1基因在酵母相和菌丝相的DNA表达水平上无统计学差异。OBJECTIVE To study the mRNA and DNA expression of PLB1 gene encoding the phospholipase B1 in the yeast form and hyphal form of Candida albicans and discuss the difference in the expression of virulence gene encoding invasive enzymes in the dimorphism of C.albicans from the perspective of molecular biology,providing a preliminary experimental basis for the further research of genetic variation,pathogenicity and prevention of C.albicans.METHODS Total RNA of C.albicans in yeast form and hyphal form were extracted by RNAiso Plus rea-gent combined with glass beads method.The differences in the mRNA expression of PLBI were observed by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR).Total DNA of C.albicans in yeast form and hyphal form were extracted separately,and the difference in DNA of PLBl gene between yeast form and hy-phal form of C.albicans were compared by polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP).RESULTS Electrophoresis analysis showed that the extracted total RNA of C.albicans in yeast form and hyphal form were three clear bands,indicating 28S rRNA,18S rRNA,and 5S rRNA,which showed the integrality of the total RNA extracted.The semi-quantitative RT-PCR showed that the relative expression of mRNA of PLB1 gene was(0.90±0.04)in yeast form and(1.00±0.01)in hyphal form,with significant difference(P<0.05).The results of SSCP map by PCR-SSCP showed that there was no significant differences in the electrophoretic bands of PLB1 gene between yeast form and hyphal form of C.albicans.CONCLUSION The concentration,purity,and integrity of total RNA extracted by RNAiso Plus reagent combined with glass beads were high.The mRNA expression level of PLB1 is significantly higher in the hyphal form than in the yeast form of C.albicans.The SSCP map obtained by PCR-SSCP showed that there is no significant difference in PLB1 DNA between the yeast form and the hyphal form of C.albicans.

关 键 词：白假丝酵母 二相性 磷脂酶B1 mRNA表达 DNA表达 

分 类 号：R379[医药卫生—病原生物学]