尖孢镰刀菌古巴专化型4号生理小种中Fsr1基因的克隆及生物信息学分析

以尖孢镰刀菌古巴专化型4号生理小种为材料,克隆其Fsr1基因的DNA及cDNA全长,并对其进行生物信息学分析。根据镰刀菌中相关基因设计简并引物,结合PCR与RT-PCR技术扩增目的片段,进行氨基酸序列推导后分析。得到结果 FoFsr1基因开放阅读框为2 474 bp,共编码824个氨基酸;FoFsr1基因编码蛋白可能是存在于细胞核中的跨膜亲水蛋白,含有54个蛋白磷酸化位点,不含有信号肽;该蛋白主要含有1个Striatin结构域及7个WD40结构域,二级结构主要由β折叠和无规则卷曲组成,其中不含有亮氨酸拉链结构;同源性分析发现其与丛赤壳菌属中的Fsr1蛋白亲缘关系最近。通过结构分析发现,推测该蛋白可能在细胞周期及细胞凋亡过程中起信号转导与转录调控的作用,从而对病原菌的致病力产生影响。

Cloning and Bioinformatics Analysis of Fsr1 Gene in Fusarium oxysporum f. sp. cubense Race 4

The objective of this study is to clone the full-length cDNA of Fsrl gene from Fusarium oxysporum f. sp. cubense race 4 and analysis Fsrl gene through bioinformatics method. We designed degenerate primers in accordance with other species Fusariums and amplified FoFsr1 gene by PCR and RT-PCR methods. The amplification products were sequenced and the amino acid sequence were aligned and analysed. Sequence analysis extrapolated that the open reading frame of this gene was 2 474 bp and encoding a putative protein of 824 amino acids. FoFsr1 protein was hydrophilic protein which likely located in the cell nuclei. This protein had likely 54 phosphorylation sites with no signal peptide. FoFsr1 protein has one Striatin and seven WD40 domain. The secondary structure was mainly constituted by the β-strand and random coil without leucine zipper. Phylogenetic tree analysis indicated that FoFsr1 protein was highly genetic relationship with Nectria haematococca. FoFsr1 protein maybe some serious functions in cell cycle and apoptosis through structural analysis and affect in pathogenic of Fusarium.