猪苓菌核2种热激蛋白基因的克隆及其表达分析  

Molecular cloning and characterization of two kind of heat-shock protein gene from Polyporus umbellatus

作　　者：刘蒙蒙[1,2] 邢咏梅[1] 郭顺星[1] 

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193 [2]江苏理工学院电气信息工程学院生物信息与医药工程研究所,江苏常州213001

出　　处：《中国中药杂志》2016年第24期4550-4555,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(31201666);2014年度山西省煤基重点科技攻关项目(FT2014-03);河北省科技计划项目(16232503D)

摘　　要：该研究采用RT-PCR技术从中国野生猪苓菌核Polyporus umbellatus中克隆得到2种热激蛋白基因,其中Pu Hsp90基因的开放阅读框2 091 bp,编码696个氨基酸,推导的蛋白质相对分子质量约78.9 k Da;Pu Hsp70基因的开放阅读框1 944 bp,编码647个氨基酸,推导的蛋白质相对分子质量约70.5 k Da;蛋白质结构预测及同源比对分析表明,这2个基因编码的核苷酸序分别具有Hsp90,Hsp70蛋白的保守结构域。进化树分析表明,Pu Hsp90与变色栓菌Hsp90聚为一类,Pu Hsp70与肝色牛排菌Hsp70聚为一类。qRT-PCR分析表明,蜜环菌侵染的情况下,这2个基因在猪苓菌核中均上调表达。这2个基因在蜜环菌侵染猪苓菌核的状态下有相同的表达模式,预示这2个基因其可能在抗生物胁迫中起重要作用。With RT-PCR approaches, the full-length cDNA of two heat shock protein genes were cloned from total RNA of the Pol- yporus umbellatus sclerotium. The full open reading frame cDNA sequence of the Hsp90 was 2 091 bp, encoding 696 amino acid resi- dues with a predicted molecular mass of 78.9 kDa. The full open reading frame cDNA sequence of the Hsp70 was 1 944 bp, encoding 647 amino acid residues with a predicted molecular mass of 70. 5 kDa. The Hsp90 and Hsp70 protein contained the conservative struc- ture domain, respectively. Phylogenetic analysis showed that Hsp90 and Hsp90 from Trametes versicolor were clustered into one group, Hsp70 and Hsp70 from Fistulina hepatica were clustered into one group. Real-time PCR analysis showed that, the expression of Hspg0 and Hsp70 in the infected part by Amillariella mellea was upregulated. The expression profiling of Hsp90 and Hsp70 showed same pat- terns underbiotic stress. The results indicate that these two genes may play an important role in response to Amillariella meUea infec- tion.

关 键 词：猪苓 热休克蛋白 基因克隆 表达分析 

分 类 号：S567.3[农业科学—中草药栽培]