药用真菌猪苓溶血素基因克隆和序列分析  

Molecular cloning and characterization of a hemolysin gene from medicinal fungus Polyporus umbellatus

作　　者：宋超[1] 郭顺星[1] 

机构地区：[1]北京协和医学院中国医学科学院药用植物研究所,北京100193

出　　处：《菌物学报》2013年第4期690-697,共8页Mycosystema

基　　金：国家自然科学基金(No.30830117);国际科技合作项目(No.2011DFA31260);高等学校博士学科点专项科研基金资助课题(No.20091106110003)

摘　　要：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 hemolysin gene relative to fungal metamorphosis was firstly cloned from medicinal fungus Polyporus umbellatus by 3＇ rapid amplification of cDNA end PCR （RACE）. A 744bp full-length cDNA of hemolysin gene contained 447bp ORF, which was predicted to encode a 334-amino acid protein of 15.79kDa with an isoelectronic point （pl） of 4.89. The deduced umbellolysin protein contained the structure domain and functional sites of aegerolysin, showing 60% identical to aegerolysin, and clustered with basidiomycete group in the phylogenetic analysis. Quantitative real-time PCR analysis revealed that umbellolysin transcripts were abundant in the beginning of sclerotial formation （20-30 days）, the transcripts levels of P. umbellatus umbellolysin gene were particularly higher in sclerotia than mycelia, suggesting that the gene was involved in sclerotial development in P umbellatus.

关 键 词：猪苓 溶血素 3'-RACE 实时荧光定量PCR 菌核 

分 类 号：S567.39[农业科学—中草药栽培]