白念珠菌菌丝相与酵母相ERG11基因PCR-RFLP比较分析

目的通过对白念珠菌菌丝相与酵母相ERG11基因限制性片段长度多态性分析,比较两相细胞在DNA水平的差异。方法分别抽提来自同一亲本、对氟康唑敏感性不同的白念珠菌(CA-1,CA-2,CA-4,CA-6,CA-7,CA-9,CA-11,CA-13~CA-17)菌丝相与酵母相基因组DNA,根据ERG11基因序列设计一对引物,对其进行PCR扩增(包括部分上游和下游非编码区),扩增产物分别用AccⅠ和MunⅠ消化、琼脂糖凝胶电泳后观察。结果各株白念珠菌菌丝相与酵母相均能扩增出约1 734bp大小的目的片段,CA-7,CA-14,CA-16和CA-17株两相细胞ERG11基因RFLP图谱存在差异,其余受试菌株未见差异。结论从DNA全貌来看,白念珠菌菌丝相与酵母相ERG11基因存在差异。

Comparative Study on The Difference of ERG11 Gene between The Yeast Form and The Hyphal Form of Candida albicans by PCR-RFLP

Objective To investigate the difference on DNA sequence between the yeast form and the hyphal form of Candida albicans,PCR-RFLP(restricted fragment length polymorphism) fingerprints of ERG11 encoding the target enzyme to fluconazole were compared.Methods The genomic DNA was extracted from the yeast or hyphal form of C.albicans(CA-1,CA-2,CA-4,CA-6,CA-7,CA-9,CA-11,CA-13~CA-17)which were isolated from a HIV-seropositive patient.One set of primes was designed to amplify ERG11 gene,and the PCR product was cleaved by restriction enzyme AccⅠ or MunⅠ,and was electrophored before oberserved.Results Target fragment of ERG11 gene(approximately 1 734 bp)was successfully amplified in all C.albicans strains,and there were differences on RFLP fingerprints between the hyphal form and the yeast form of CA-7,CA-14,CA-16 and CA-17 C.albicans strains.Conclusion In general,there are significant differences on ERG11 gene sequences between the hyphal form and the yeast form of C.albicans.