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目的克隆犬小孢子菌丝氨酸水解酶FSH1基因全长c DNA,为进一步研究FSH1的功能以及其在头癣中的致病机理打下基础。方法以犬小孢子菌的总RNA为材料,采用c DNA末端快速扩增(Rapid c DNA end amplification,RACE)方法扩增犬小孢子菌丝氨酸水解酶FSH1基因的全长c DNA。结合生物学软件对其序列进行了生物信息学分析。结果该全长c DNA大小为945bp,包含44bp的5’UTR、70bp的3’UTR、开放阅读框为831bp,编码276个氨基酸。Blast搜索结果显示,该全长c DNA与已报道的编码犬小孢子菌的结构域蛋白的基因序列同源性达100%,与编码石膏样小孢子菌的结构域蛋白基因序列同源性达78%。结论首次成功获得犬小孢子菌丝氨酸水解酶FSH1基因的全长c DNA,为进一步研究其在头癣发病中的功能奠定了基础。
Objective To clone full-length cDNA of FSH1, a serine hydrolase of microsporum, to lay the foundation for further study on the function of FSH1 and its pathogenesis in tinea capitis. Methods The total RNA of FSH1 gene of microsporum could be amplified by rapid c DNA end amplification (RACE) using total RNA of Microsporum canis as the material. Bioinformatics analysis of the sequence was carried out with biological software. Results The full-length c DNA was 945bp in length and contained 44bp 5’UTR and 70bp 3’UTR. The open reading frame was 831bp encoding a protein of 276 amino acids. Blast search results showed that the full-length cDNA has a homology of 100% with the reported domain protein of Microsporum canis, which is homologous to the sequence of the domain protein encoding Microsporum gypseum 78%. Conclusion The full-length c DNA of serine hydrolase FSH1 gene of Microsporum canis was successfully obtained for the first time, which laid the foundation for further study on its function in the pathogenesis of tinea capitis.