烟曲霉新基因pim1敲除菌株构建及其耐药功能鉴定

Construction of a New Gene pim1 Knockout Strain of Aspergillus Fumigatus and Identification of Its Drug Resistance Function

在前期构建的T-DNA(Transfer DNA)随机插入的烟曲霉耐唑类药物突变菌株的基础上,通过Tail-PCR和测序,从突变菌株中鉴定到一个新基因pim1,并进一步利用基因敲除技术构建烟曲霉pim1基因敲除菌株,从而通过菌落表型验证该基因缺失所导致的耐药现象.由此推测,pim1基因可能通过影响烟曲霉线粒体的功能参与唑类药物耐药的调控.该研究结果可为后期深入研究pim1基因在烟曲霉中参与唑类药物耐药分子调控机制,以及进一步控制真菌的耐药现象提供理论依据.

In this study, a new gene pim1 was identified from the mutant strain by Tail-PCR and sequencing on the basis of the T-DNA(Transfer DNA) randomly inserted into the azole-resistant mutant strain of Aspergillus fumigatus, and the pim1 gene knockout strain of the Aspergillus fumigatus was further constructed by gene knockout technology, and the drug resistance caused by the deletion of the gene was verified by the colony phenotype. We speculate that the pim1 gene may be involved in the regulation of azole drug resistance by affecting the mitochondrial function of Aspergillus fumigatus. The results of this study can provide a theoretical basis for the in-depth study of pim1 gene in Aspergillus fumigatus in the molecular regulation mechanism of azole drug resistance and the control of fungal resistance.