Efficient insertional mutagenesis system for the dimorphic pathogenic fungus Sporothrix schenckii using Agrobacterium tumefaciens
Efficient insertional mutagenesis system for the dimorphic pathogenic fungus Sporothrix schenckii using Agrobacterium tumefaciens
复制标题
利用根癌农杆菌对二形性致病真菌申克孢子丝菌进行高效插入诱变系统
DOI：
10.1016/j.mimet.2011.01.017
复制
发表时间：
2011-03-01
影响因子：
2.2
通讯作者：
Wang, Li
中科院分区：
文献类型：
--
作者：
Zhang, Yanhua;Li, Guangquan;Wang, Li
申克孢子丝菌是一种在全球范围内引起人畜孢子虫病的二型致病真菌。本研究建立并优化了根癌农杆菌介导的转化体系。Schenckii用于插入诱变。转化效率达到每10（6）个分生孢子600个以上转化子。使用该方案使我们能够在短的实验时间内获得大量的T-DNA插入突变体。在转化实验中获得了几个表型改变的突变体。突变体显示出有丝分裂稳定性。通过热不对称交错PCR（TAIL-PCR）克隆转移DNA（T-DNA）侧翼序列。我们的结果表明，ATMT系统可以作为一个有效的工具，在S。申克。这是第一次报告的一个合适的诱变系统，可能提供有价值的突变体和信息，在未来的正向和反向遗传学研究，这种医学上重要的真菌。(C)2011爱思唯尔有限公司版权所有。
Sporothrix schenckii is a dimorphic pathogenic fungus that causes human and animal sporotrichosis globally. Here we developed and optimized an Agrobacterium tumefaciens-mediated transformation (ATMT) system of S. schenckii for insertional mutagenesis. The transformation efficiency reached more than 600 transformants per 10(6) conidia. Using this protocol enabled us to obtain a large number of T-DNA insertional mutants within a short experimental period. Several mutants with altered phenotypes were obtained during the transformation experiments. The mutants displayed mitotic stability. Transferred DNA (T-DNA) flanking sequences were cloned by thermal asymmetric interlaced PCR (TAIL-PCR). Our results demonstrated that the ATMT system can be an effective tool for insertional mutagenesis in S. schenckii. This is the first report of a suitable mutagenesis system which may provide valuable mutants and information for both forward and reverse genetics research in the future for this medically important fungus. (C) 2011 Elsevier B.V. All rights reserved.