根瘤农杆菌T-DNA插入介导的烟曲霉耐药突变菌的分子机制  

Molecular mechanism of Agribacterium tumefaciens T-DNA insertion mediated Aspergillus fumigatus drug resistant mutant

作　　者：龙南彪 LONG Nanbiao(School of Medical Technology, Shaoyang University, Shaoyang 422000, China)

机构地区：[1]邵阳学院医学技术学院,湖南邵阳422000

出　　处：《邵阳学院学报（自然科学版）》2021年第5期1-7,共7页Journal of Shaoyang University：Natural Science Edition

基　　金：湖南省教育厅优秀青年项目(18B434);湖南省自然科学基金青年项目(2019JJ50562);国家自然科学基金青年项目(32000107)。

摘　　要：目的探讨一株根瘤农杆菌T-DNA插入介导的烟曲霉突变菌(M7)耐药性产生的分子机制。方法通过基因组重测序鉴定M7中T-DNA的插入位置并分析SNP位点,然后在烟曲霉A1160背景上将相应的突变基因进行敲除并验证。敲除成功后,比较分析敲除菌和野生型菌株的表型。结果通过基因组重测序发现M7中T-DNA的插入位点位于基因Afu5g08910中,但其敲除菌(ΔM7)并未发现有耐药性产生。SNP分析以及验证实验发现,M7中Cyp51A第54位氨基酸由Gly突变成Arg是M7产生耐药性的根本原因。结论M7突变株耐药性产生的原因并非Afu5g08910基因突变,而是Cyp51A位点突变所致。Objective To investigate the molecular mechanism of Aspergillus fumigatus mutant(M7)to azolesre sistance that mediated by Agrobacterium tumefaciens T-DNA insertion.Methods The insertion sites of T-DNA in M7 mutant strains were identified by genome resequencing and SNP sites were also analyzed.Next,the possibly related mutant genes were knocked out and verified by PCR in the background of Aspergillus fumigatus A1160.The phenotypes of knockout and wild-type strains were compared and analyzed.Results The T-DNA insertion site in M7 was found to be located in gene Afu5g08910 by genome re-sequencing,which was verified by diagnostic PCR.However,null mutant of the Afu5g08910 gene(ΔM7)did not reveal resistance to anti-fungal drugs.SNP analysis revealed that the 54th position of Cyp51A were mutated from Gly to Arg,which was the primary cause of drug resistance of M7.Conclusion The drug resistance of M7 mutant is not caused by mutation of Afu5g08910 gene but due to mutation of Cyp51A.

关 键 词：烟曲霉 T-DNA 耐药性 

分 类 号：R379.6[医药卫生—病原生物学]