基于PCR-LIS-SSCP的白念珠菌菌丝相与酵母相ERG11基因比较研究

A comparative study on the difference in the sequence of ERG11 gene between the yeast and hyphal form of Candida albicans by PCR-LIS-SSCP

目的通过对白念珠菌菌丝相与酵母相ERG11基因PCR-LIS-SSCP图谱的比较,探讨两相细胞在DNA水平的差异。方法分别抽提16株来自同一亲本且对氟康唑敏感性不同的白念珠菌菌丝相与酵母相基因组DNA,根据ERG11基因序列设计7对引物,对其进行分段PCR扩增,扩增产物经单链变性后,以非变性聚丙烯酰胺凝胶进行SSCP分析。结果16株白念珠菌菌丝相与酵母相均能扩增出目的片段,SSCP图谱显示两相细胞的ERG11基因序列存在多位点差异。结论白念珠菌菌丝相与酵母相ERG11,基因存在着差异。

Objective To investigate the difference of ERG11 gene between the yeast and hyphal forms of Candida albicans by PCR-LIS-SSCP （ low ionic strength single-strand conformation polymorphism ）. Methods The genome DNA was extracted from the yeast and hyphal forms of 16 strains of C. albicans, which were isolated from a HIV-seropositive patient and showed different susceptibility to fluoconazole. ERGII gene was amplified by PCR with seven sets of primers. Then the PCR products were denatured and subjected to PCR-LIS-SSCP analysis. Results Seven fragments of the ERGI 1 gene were successfully amplified for both forms of all 16 strains. As shown by PCR-LIS-SSCP fingerprints, differences were observed in many sites of ERG11 gene between the two forms of C. albicans. Conclusion In general, differences exist in the ERG11 gene sequence between the hyphal form and yeast form of C.albicans.