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利用3’-RACE-PCR方法首次从药用真菌猪苓中克隆得到与真菌形态发育相关的溶血素基因。结果表明,猪苓溶血素基因的全长cDNA为744bp,其中编码区占447bp,共编码148个氨基酸,推测其分子量约为15.79kDa,理论等电点为4.89。推定的猪苓溶血素蛋白具有与杨树菇溶血素类蛋白家族相同的结构域和功能位点,两者同源性为60%。系统进化树结果显示猪苓溶血素隶属于担子菌类群。实时荧光定量PCR分析结果表明在菌核形成初期猪苓溶血素基因表达量较高,且显著高于菌丝体中猪苓溶血素基因的转录水平,说明溶血素基因参与了猪苓菌核的形态发育。
The 3’-RACE-PCR method was firstly used to clone the hemolysin gene related to fungal morphological development from the medicinal fungus Polyporus umbellatus. The results showed that the full length cDNA of uralin was 744bp in length, encoding 447bp of which encoded 148 amino acids. The deduced molecular weight was about 15.79 kDa and the theoretical isoelectric point was 4.89. The putative urinolysin has the same domain and function as the poplar glucosinolate family, with a homology of 60%. Phylogenetic tree results show that the polyrin hemolysin belongs to the basidiomycetes group. The results of real-time PCR showed that the expression of PPL gene was higher in the early stages of sclerotia and was significantly higher than that in the mycelium, indicating that the hemolysin gene is involved in the Morphological development.