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Efficient gene replacements in ku70 disruption strain of Aspergillus chevalieri var. intermedius

文献类型： 外文期刊

作者： Huang, Qingqing 1 ; Cao, Yang 2 ; Liu, Zuoyi 2 ; Tan, Yumei 2 ; Liu, Yongxiang 3 ;

作者机构： 1.Guizhou Normal Univ, Dept Life Sci, Guiyang, Peoples R China

2.Guizhou Acad Agr Sci, Guizhou Key Lab Agr Biotechnol, Guiyang, Peoples R China

3.Guizhou Acad Agr Sci, Guizhou Inst Biotechnol, Guiyang, Peoples R China

关键词： Aspergillus chevalieri var. intermedius;ku70;gene replacement;homologous recombination;nonhomologous end-joining

期刊名称：BIOTECHNOLOGY & BIOTECHNOLOGICAL EQUIPMENT （ 影响因子：1.632； 五年影响因子：2.029 ）

ISSN： 1310-2818

年卷期： 2017 年 31 卷 1 期

页码：

收录情况： SCI

摘要： Aspergillus chevalieri var. intermedius is a dominant filamentous fungal species in Fuzhuan tea and is associated with the quality and health benefits of this tea. The sexual or asexual reproduction of this fungus depends on the osmotic pressure of the tea. Efforts to enhance the beneficial effects of A. chevalieri var. intermedius are hampered by difficulties in disrupting its genes. To address this issue, we identified the A. chevalieri var. intermedius homolog (Acku70) of human Ku70 and generated an Acku70 disruption strain (Delta Acku70), aiming to improve the gene replacement efficiency. Delta Acku70 grew at a slightly lower rate in vitro than the wild-type strain; however, the two strains exhibited similar sensitivity to temperature, osmotic pressure and the effects of ethyl methane sulphonate and H2O2. The replacement efficiency of veA and flbA dramatically increased in Delta Acku70 compared to that in the wild type. The efficiency of flbA replacement increased from 2.6% to 80%, whereas the frequency of veA disruption increased from 15.2% to 83.9% and from 30.8% to 86.8%. Thus, Delta Acku70 is suitable for use as a type strain for large-scale functional genomic analysis of A. chevalieri var. intermedius.

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