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Effect of heterologous auxotrophy markers on adaptation to various stresses in Candida albicans
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- Author:
- No author available
- Journal Title:
- Academic Journal of Second Military Medical University
- Issue:
- 9
- DOI:
- 10.3724/SP.J.1008.2012.00929
- Key Word:
- 白假丝酵母菌;基因敲除;异源性筛选标记;应激
Abstract： 目的 考察以His1-Leu2-Arg4基因敲除策略敲除白假丝酵母菌基因后,异源性筛选标记C.dubliniensis HIS1(C.d.HIS1)、C.maltosa LEU2(C.m.LEU2)和C.dubliniensis ARG4(C.d.ARG4)对菌株耐受各种应激的影响.方法 以SN152为亲本菌,融合PCR法构建重组片段,醋酸锂法转染白假丝酵母菌SN152,分别构建上述3个标记的回复菌.采用套式PCR法鉴定回复菌的构建,spot assay实验考察菌株对各种应激的敏感性.结果 PCR验证7种菌株构建成功,spot assay结果显示各回复菌对pH应激、氧化应激、各种临床抗菌药应激、金属离子应激、细胞壁损伤相关药物应激、渗透应激、DNA损伤化合物应激的敏感性与亲本菌SN152一致.而给予0.02％ SDS时各菌株的敏感性不一致,缺乏C.d.HIS1标记的菌株在0.02％SDS的应激条件下不能生长.结论 C.d.HIS1、C.m.LEU2和C.d.ARG4 3个异源性筛选标记不影响白假丝酵母菌对大部分应激的敏感性,不同标记基因敲除菌间可以相互比较,且可以统一用SN152作为对照菌株.在考察SDS应激时,所用筛选标记不同,对结果有影响,不能用SN152代替作为统一对照,若要相互比较,要保证C.d.HIS1标记必须一致.
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