烟曲霉突变体库的构建及伊曲康唑耐药菌的筛选

中国真菌学杂志　 2020, Vol. 15 　Issue (1): 47-51.

烟曲霉突变体库的构建及伊曲康唑耐药菌的筛选

刘晶, 徐稳, 谭志伟, 刘瑶瑶, 周玲玲, 付裕, 龙南彪

邵阳学院医学检验学院, 邵阳 422000

收稿日期:2019-04-23 出版日期:2020-02-28 发布日期:2020-02-28
通讯作者: 龙南彪,E-mail:2008-lnb@163.com E-mail:2008-lnb@163.com
作者简介:刘晶,女(汉族),本科,职称.E-mail:1652859919@qq.com
基金资助:
湖南省教育厅优秀青年项目（18B434）；湖南省自然科学基金（2019JJ50562）；湖南省大学生研究性学习和创新性实验计划项目（201810547024）

The construction of Aspergillus fumigatus mutant library and screening of itraconazole drug-resistant mutant

LIU Jing, XU Wen, TAN Zhi-wei, LIU Yao-yao, ZHOU Ling-ling, FU Yu, LONG Nan-biao

Department of Laboratory Medicine, Shaoyang University, Shaoyang 422000, China

Received:2019-04-23 Online:2020-02-28 Published:2020-02-28

摘要/Abstract

摘要：

目的构建烟曲霉突变体库并筛选耐药突变子，初步探讨烟曲霉的耐药机制。方法利用根瘤农杆菌T-DNA介导的插入突变构建烟曲霉突变体库，用伊曲康唑对突变体库进行筛选，挑取烟曲霉耐药突变株。结果利用根瘤农杆菌T-DNA插入突变，本研究获得了1805株突变子，诊断PCR显示突变子基因组DNA上均含有潮霉素标记，说明T-DNA均插入到了宿主的基因组上。通过筛选，本研究成功获得了16株烟曲霉伊曲康唑耐药菌，其中有两株对两性霉素B表现为不同程度的耐药性。结论通过构建烟曲霉突变体库以及耐药突变子筛选获得了16株烟曲霉耐药菌，为进一步研究烟曲霉的耐药机制奠定了基础。

关键词: 烟曲霉, 侵袭性曲霉病, T-DNA, 耐药性

Abstract:

Objective This study aim to construct a A. fumigatus mutant library and to screen the drug resistance mutant for preliminarily investigation of the mechanism of A. fumigatus drug resistance. Methods The A. fumigatus mutant library was constructed by Agrobacterium tumefaciens T-DNA insertion. For drug resistance mutant of A. fumigatus, plates that containing anti-fungal drug itraconazole were used. Results The present study obtained 1805 A. fumigatus mutants. Diagnostic PCR result showed that each tested mutant had a hygromycin marker, indicating that T-DNA were inserted into the host genome. Further, 16 strains of A. fumigatus mutants were successfully obtained by screening the mutant library using itraconazole, among which 2 exhibited obvious Amphotericin B resistance. Conclusion Sixteen A. fumigatus drug-resistant strains were obtained by construct and screening the A. fumigatus mutant library, which laid a foundation for further study on the mechanism of drug resistance of A. fumigatus.

Key words: Aspergillus fumigatus, invasive aspergillosis, T-DNA, drug resistance

中图分类号:

R379.6

刘晶, 徐稳, 谭志伟, 刘瑶瑶, 周玲玲, 付裕, 龙南彪. 烟曲霉突变体库的构建及伊曲康唑耐药菌的筛选[J]. 中国真菌学杂志, 2020, 15(1): 47-51.

LIU Jing, XU Wen, TAN Zhi-wei, LIU Yao-yao, ZHOU Ling-ling, FU Yu, LONG Nan-biao. The construction of Aspergillus fumigatus mutant library and screening of itraconazole drug-resistant mutant[J]. Chinese Journal of Mycology, 2020, 15(1): 47-51.

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