白念珠菌FLO8基因高表达菌株构建及鉴定  

Construction and identification of FLO8-overexpressing strain of Candida albicans

作　　者：李文静[1,2] 刘锦燕[2] 史册[1] 王影[1,2] 赵悦[1,2] 项明洁[1,2] 

机构地区：[1]上海交通大学医学院附属瑞金医院检验科,上海200025 [2]上海交通大学医学院附属瑞金医院卢湾分院放免检验科,上海200020

出　　处：《诊断学理论与实践》2016年第1期9-14,共6页Journal of Diagnostics Concepts & Practice

基　　金：上海市科委基金(15ZR1426900);上海市医学重点专科(ZK2012A21);上海市黄浦区优秀青年人才(RCPY1407)

摘　　要：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 FLO8-overexpressing strain of Candida albicans. Methods Candida albicans FLO8 gene was inserted into ADH1 promoter in p CP20 plasmid vector. Then the fragment of ADH1-FLO8-Lox P-ARG4-Lox P-ADE2 obtained by PCR was integrated into SN152 strain at the endogenous ADE2 loci by homologous recombination. Results FLO8-overexprssing plasmid was constructed as confirmed by DNA sequencing. FLO8 gene was integrated into the ADE2 gene loci of Candida albicans SN152 and expressed highly as identified by homologous recombination and real-time reverse transcription-PCR. Conclusions FLO8-overexpressing strain of Candida albicans can be efficiently constructed with p CP20 plasmid as vector by using techniques of homologous recombination.

关 键 词：白念珠菌 FLO8基因 高表达 

分 类 号：R519.5[医药卫生—内科学]