烟曲霉组氨酸激酶基因片段的隆及表达分析

Cloning and Expression of Histidine Kinase Gene of Aspergillus Fumigatus

目的分离克隆烟曲霉组氨酸激酶基因,进一步探讨其在侵袭性感染中的作用。方法基于组氨酸激酶保守区设计简并引物,通过逆转录-聚合酶链反应获得特异性扩增产物,对目的产物克隆、测序。通过Blast分析进行相似性比较。以获得的目的基因片段作为探针,通过Northern杂交比较该基因在体外与体内感染时表达的差异。结果逆转录-聚合酶链反应获得2个片段,长度分别为171bp、305bp,其中305bp片段与构巢曲霉tesA基因的同源性>80%。该基因在小鼠侵袭性肺曲霉病中表达水平明显增加。结论烟曲霉组氨酸激酶基因的表达可能有助于其在体内存活,该基因可能是烟曲霉潜在的毒力基因。

Objective Through isolating the histidine kin ase gene of Aspergillus fumigatus,to detect its role in the invasive aspergillosis.Methods Using degenerate primers for highly conserved regions of his-tidine kinase,RT -PCR was performed with cDNA of Aspergillus fumigatus as a template.The gene expression of it in vitro and in vivo was investig ated by Northern blot.Results A fragment of this gene was cloned fro m Aspergillus fumigatus that is highly homologous to tesA gene of Aspergillus nidulans,which was expressed at high level during invasive infection.Conclusion The results indicate that this gene may attribute to the invasive aspergillosis.