扁桃酸代谢对新生隐球菌毒力作用的探讨

发布时间：2018-04-16 08:10

本文选题：新生隐球菌 + 扁桃酸　；参考：《中国真菌学杂志》2015年06期

【摘要】：目的探讨扁桃酸代谢对新生隐球菌毒力作用的影响。方法采用套叠PCR方法,构建扁桃酸消旋酶、扁桃酸脱氢酶及过氧化氢铜胺氧化酶基因重组片段,利用基因枪将重组片段转入新生隐球菌JEC21,应用PCR筛选、DNA测序等方法对阳性克隆子进行筛选与鉴定。体外观察并检测突变菌株对多种应激条件的应答反应。结果成功构建了新生隐球菌JEC21扁桃酸消旋酶、扁桃酸脱氢酶及过氧化氢铜胺氧化酶的基因缺陷菌株(mreΔ,lmdΔ,pcaoΔ),发现缺陷菌株mreΔ对于毒力功能影响不大,pcaoΔ、lmdΔ在高温、高渗、氧化应激等方面毒力减退,而黑色素分泌增加。结论扁桃酸代谢参与介导新生隐球菌毒力作用的调控,但其具体机制有待于进一步的研究探讨。
[Abstract]:Objective to investigate the effect of mandelic acid metabolism on virulence of Cryptococcus neoformans.Methods the recombinant gene fragments of mandelic acid racemase, mandelic dehydrogenase and copper amine oxidase were constructed by overlapping PCR.The recombinant fragment was transferred into Cryptococcus neoformans JEC21 by gene gun. The positive clones were screened and identified by PCR screening and sequencing.The response of mutant strains to various stress conditions was observed and tested in vitro.Results the gene deficient strains of Cryptococcus neoformans JEC21 mandelic acid racemase, mandelic dehydrogenase and copper amine oxidase hydrogen peroxide were successfully constructed. It was found that the defective strain mre 螖 had little effect on virulence.Oxidative stress and other aspects of toxicity decreased, while the secretion of melanin increased.Conclusion mandelic acid metabolism is involved in the regulation of Cryptococcus neoformans virulence, but its specific mechanism needs further study.
【作者单位】：上海市医学真菌分子生物学重点实验室第二军医大学附属长征医院皮肤科;河南省濮阳市油田总医院皮肤科;
【基金】：国家973项目(2013CB531601) 国家自然科学基金(31270180) 上海市医学真菌分子生物学实验室基金(14DZ2272900)
【分类号】：R379.5

【共引文献】