猪苓菌核2种热激蛋白基因的克隆及其表达分析

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。

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

With RT-PCR approaches, the full-length cDNA of two heat shock protein genes were cloned from total RNA of the Polyporus umbellatus sclerotium. The full open reading frame cDNA sequence of the Hsp90 was 2 091 bp, encoding 696 amino acid residues 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 structure 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 Hsp90 and Hsp70 in the infected part by Amillariella mellea was upregulated. The expression profiling of Hsp90 and Hsp70 showed same patterns underbiotic stress. The results indicate that these two genes may play an important role in response to Amillariella mellea infection.