冠突曲霉产孢相关基因的克隆及功能分析

发布时间：2018-02-25 05:29

本文关键词： 冠突曲霉有性发育 Tail-PCR NDT　出处：《基因组学与应用生物学》2017年09期 　论文类型：期刊论文

【摘要】：为探究冠突曲霉有性产孢调控的相关机制,从已构建的T-DNA随机插入突变体库中选取有性产孢突变株,采用Southern杂交鉴定T-DNA为单拷贝插入,并用Tail-PCR技术扩增获得T-DNA插入位点侧翼序列,分离获得突变基因。序列分析表明,该基因位于冠突曲霉基因组scaffold 7上,开放阅读框2 511 bp,含有4个内含子,预测编码666个氨基酸,含有NDT80_Pho G蛋白质超级家族保守结构域。BLASTp结果表明,该预测蛋白与Eurotium rubrum p53-like转录因子(EYE95652.1)相似性最高,identity=89%。显微观察发现T-DNA的插入影响了有性产孢结构的发育。本实验为进一步研究该基因在有性产孢过程中的分子功能及可能参与的调控途径奠定基础。
[Abstract]:In order to explore the mechanism of regulation of sexual sporulation in Aspergillus coronatum, a single copy of T-DNA was identified by Southern hybridization from the constructed T-DNA random insertion mutant library. The flanking sequence of T-DNA insertion site was amplified by Tail-PCR, and the mutant gene was isolated. The sequence analysis showed that the gene was located on the genomic scaffold 7 of Aspergillus coronatum, open reading frame 2511 BP, containing 4 introns and encoding 666 amino acids. The conserved domain of superfamily containing NDT80_Pho G protein. BLASTp showed that, The predicted protein has the highest similarity with Eurotium rubrum p53-like transcription factor EYE95652.1). Microscopic observation shows that the insertion of T-DNA affects the development of sexual sporulation structure. In order to further study the molecular function of this gene in the process of sexual sporulation. To be able to participate in the regulation of the way to lay the foundation.
【作者单位】：贵州大学;贵州省生物技术研究所;贵州省农业生物技术重点实验室;贵州省农业科学院;
【基金】：国家自然科学基金“冠突曲霉有性产孢新基因的筛选及其功能验证”(31660021) 贵州省农业科学院自主创新专项“冠突散囊菌比较基因组学及功能基因组学研究”(2014)004 贵州省农科院与贵州省科技厅联合基金项目(黔科合LH字(2014)7693号) 贵州大学研究生创新基金“冠突散囊菌产孢相关基因的克隆及功能分析”共同资助
【分类号】：Q78;Q93

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