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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。
Fusarium oxysporum Cubaspecific 4 race was used as material to clone the full-length cDNA and cDNA of Fsr1 gene, and its bioinformatics analysis was carried out. The degenerate primers were designed according to the related genes in Fusarium and the target fragments were amplified by PCR and RT-PCR. The amino acid sequences were deduced and analyzed. The results showed that the open reading frame of FoFsr1 gene was 2 474 bp, encoding a total of 824 amino acids. The FoFsr1 gene encoding protein may be a transmembrane hydrophilic protein present in the nucleus, containing 54 protein phosphorylation sites without signal peptide; The protein mainly contains one Striatin domain and seven WD40 domains. The secondary structure mainly consists of β-sheet and random coil, which contains no leucine zipper structure. The homology analysis revealed that Fsr1 protein is most closely related. Through structural analysis, we found that the protein may play a role in signal transduction and transcriptional regulation during the cell cycle and apoptosis, and thus affect the pathogenicity of pathogenic bacteria.