| 摘要: |
| 目的 研究白假丝酵母菌液泡相关基因的功能，寻找白假丝酵母菌潜在的药物靶点。方法 选取 ORF19.2734、ORF19.6950、ORF19.6605、ORF19.2006 和 ORF19.5780 等5个潜在液泡功能相关基因，通过同源重组的方法构建基因缺失菌。分别观察这些基因缺失菌酵母态及菌丝态生长、液泡形态以及对各种刺激的敏感性，在阳性表型的基础上进一步研究基因功能。结果 ORF19.6950、ORF19.6605、ORF19.2006 和 ORF19.5780 基因缺失菌在酵母态及菌丝态生长、液泡形态等方面与野生菌SN152无明显差异。ORF19.2734基因缺失菌酵母态生长速率减慢，对酮康唑敏感性升高，但其液泡形态未见明显改变。RT-PCR显示，ORF19.2734基因缺失菌与野生菌SN152相比，MDR1、ERG11、ERG2 基因表达量降低。结论 ORF19.2734基因通过升高 MDR1、ERG11、ERG2 基因表达，降低菌株对唑类药物的敏感性。ORF19.2734基因可能是抗白假丝酵母菌耐药性产生的潜在靶点。 |
| 关键词: 白假丝酵母菌 液泡 ORF19.2734 基因缺失 真菌抗药性 |
| DOI：10.3724/SP.J.1008.2014.00929 |
| 投稿时间：2014-01-11修订日期：2014-03-25 |
| 基金项目:国家自然科学基金青年基金（31000079，81330083），国家重点基础研究发展计划（“973”计划，2013CB531602）. |
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| ORF19.2734 and other genes involved in vacuole biogenesis in Candida albicans：a study of functions |
| KANG Ye△,ZHOU Mi△,YAN Lan*,JIANG Yuan-ying* |
(New Drug Research and Development Center, School of Pharmacy, Second Military Medical University, Shanghai 200433, China
△Co-first authors.
* Corresponding author.) |
| Abstract: |
| Objective To study the genes involved in vacuole biogenesis in Candida albicans, so as to identify potential targets for antifungal drugs. Methods Five genes with potential involvement in vacuole biogenesis, ORF19.2734, ORF19.6950, ORF19.6605, ORF19.2006, and ORF19.5780, were selected for this study. The disruption of these target genes in Candida albicans was achieved by homologous recombination. Then the growth rate, hyphal formation, sensitivity to stresses, and the vacuole morphology of the mutants were analyzed. Results The disruptions of ORF19.6950, ORF19.6605, ORF19.2006, or ORF19.5780 resulted in no difference from the wild type strain in terms of growth rate, hyphal formation, sensitivity to stresses, and the vacuole morphology. The disruption of ORF19.2734 led to slower growth and higher sensitivity to ketoconazole, but showed no noticeable changes in vacuole mophogenesis. RT-PCR analysis found that mRNA expression of MDR1, ERG11, and ERG2 in the null mutant of ORF19.2734 was decreased compared with the wild type strain. Conclusion The null mutant of ORF19.2734 down-regulates the expression of MDR1, ERG11, and ERG2, therefore contributing to the increased sensitivity to azoles. Our results suggest that ORF19.2734 may be a potential treatment target for drug resistance in C. albicans. |
| Key words: Candida albicans vacuole ORF19.2734 gene deletion fungal drug resistance |
