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.
出处
《中国中药杂志》
CAS
CSCD
北大核心
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)
关键词
猪苓
热休克蛋白
基因克隆
表达分析
Polyporus umbellatus
heat shock protein
gene clone
gene expression