烟曲霉Bem46基因敲除株构建及其功能初步研究

发布时间：2018-01-08 00:07

本文关键词：烟曲霉Bem46基因敲除株构建及其功能初步研究　出处：《中国人兽共患病学报》2017年08期 　论文类型：期刊论文

更多相关文章： Bem 烟曲霉 发芽率 信号传导

【摘要】：目的构建烟曲霉Bem46基因敲除株,初步明确Bem46基因在烟曲霉生长出芽和各种压力传导中的作用,以及对细胞壁形成的影响。方法采用生物信息学方法查找烟曲霉中Bem46基因,设计引物,并以pyrG作为筛选标记构建烟曲霉Bem46基因敲除株(ΔBem46)。观察含不同压力物质培养基上对照菌株和ΔBem46径向生长速度,观察两菌株发芽状况以及在含有细胞壁抑制剂培养基上的生长状况。结果通过序列比对在烟曲霉基因组中找到了Bem46基因,其编号为Afu7g04660,由1 116bp碱基组成,编码311个氨基酸。使用原生质体法得到烟曲霉ΔBem46并经PCR和Southernblot确认。经观察在含有山梨醇作为渗透压力来源的培养基上ΔBem46生长明显较对照菌株快。显微镜下观察到在GMM液体培养基中,ΔBem46发芽速度慢于对照株。结论烟曲霉Bem46基因涉及由山梨醇引起的渗透压压力传导,并且该基因对促进孢子发芽可能有作用。
[Abstract]:Objective to construct the Bem46 knockout plant of Aspergillus fumigatus and to clarify the role of Bem46 gene in the growth of Aspergillus fumigatus and various pressure conduction. Methods the Bem46 gene in Aspergillus fumigatus was searched by bioinformatics and primers were designed. PyrG was used as the screening marker to construct the Bem46 knockout plant (螖 Bem46) of Aspergillus fumigatus. The radial growth rate of the control strain and 螖 Bem46 on the medium containing different pressure substances was observed. Results the Bem46 gene was found in the genome of Aspergillus fumigatus by sequence alignment. Its number is Afu7g04660, which is composed of 1116 BP base. The 螖 Bem46 of Aspergillus fumigatus was obtained by protoplast method and confirmed by PCR and Southernblot. The medium containing sorbitol as the source of osmotic pressure was observed. The growth of 螖 Bem46 was significantly faster than that of the control strain. It was observed in GMM liquid medium under microscope. Conclusion the Bem46 gene of Aspergillus fumigatus is involved in osmotic pressure conduction induced by sorbitol, and this gene may play an important role in promoting spores germination.
【作者单位】：山西医科大学第二医院皮肤科;
【基金】：国家青年科学基金项目资助(No.81101233) 山西省青年科技研究基金(No.2010021036-2) 山西省卫生计生委科研课题(No.2015043) 山西省回国留学人员科研资助项目(No.2016-052)联合资助~~
【分类号】：R379
【正文快照】： 烟曲霉是自然界常见的一种丝状真菌,在正常人群并不会由于经常暴露于烟曲霉而发生感染。然而近年来随着新技术的开展和免疫抑制剂的应用,免疫力低下的患者往往由于暴露于烟曲霉而引起侵袭性曲霉病的发生,其发病率在临床呈现明显上升趋势且病死率极高,因此探讨烟曲霉感染的发病

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