犬小孢子菌丝氨酸水解酶FSH1全长cDNA的克隆及序列分析

Cloning and Sequence Analysis of the Full-length cDNA of Microsporum Canis Serine Hydrolases( FSH1) Gene

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 Full-length cDNA of microsporum canis serine hydrolases （ FSH1 ） gene was cloned and sequenced in order to provide the basement of the further investigation of the role of FSH1 and the pathogenesis in tinea capitis. Methods Total RNA was extracted from the microsporum canis strain. The full-length cDNA sequence of FSH1 gene was cloned from total RNA as template by Rapid cDNA end amplification （RACE） method. Then it was analyzed by bioinformatics software. Results The full-length cDNA of FSH1 gene was 945bp, including a 44bp 5＇ untranslated region （URT） , a 70bp 3＇ URT and a 831bp of open reading frame （ORF） , which encoded protein of 276 amino acids. It was found by BLAST analysis that the cloned cDNA sequence was 100% and 79% homologous to those of encoding the microsporum canis and arthroderma gypse- um domain-containing protein, respectively. Conclusions The full length cDNA of microsporum canis FSH1 gene was successfully cloned for the first time. It proposed a good foundation for further investigations of the role of FSH1 on tinea capitis infected by microsporum canis.