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目的 :构建白念珠菌FLO8基因高表达菌株。方法 :将白念珠菌FLO8基因插入p CP20质粒载体ADH1启动子后,通过聚合酶链反应(polymerase chain reaction,PCR)将ADH1-FLO8-Lox P-ARG4-Lox P-ADE2片段扩增,并通过同源重组技术将该片段整合至SN152的ADE2位点。结果:通过测序鉴定FLO8基因高表达质粒载体构建成功;通过同源重组及实时反转录PCR验证表明FLO8基因整合到SN152菌株的ADE2位点并且表达量增高。结论:以p CP20质粒为载体,通过同源重组等技术,可高效构建白念珠菌FLO8基因高表达菌株。
Objective: To construct a high-expression strain of Candida albicans FLO8 gene. Methods: The Candida albicans FLO8 gene was inserted into the pCP20 plasmid vector ADH1 promoter. The ADH1-FLO8-Lox P-ARG4-Lox P-ADE2 fragment was amplified by polymerase chain reaction (PCR) Homologous recombination technique integrated this fragment into the ADE2 site of SN152. Results: The FLO8 gene expression plasmid vector was successfully constructed by sequencing. The homologous recombination and real-time reverse transcriptase-PCR confirmed that the FLO8 gene was integrated into the ADE2 locus of SN152 strain and the expression level was increased. CONCLUSION: The high expression of FLO8 gene of Candida albicans can be efficiently constructed by using pCP20 plasmid as vector and by homologous recombination.