An efficient tool for random insertional mutagenesis: Agrobacterium tumefaciens-mediated transformation of the filamentous fungus Aspergillus terreus
An efficient tool for random insertional mutagenesis: Agrobacterium tumefaciens-mediated transformation of the filamentous fungus Aspergillus terreus
复制标题
随机插入诱变的有效工具：根癌农杆菌介导的丝状真菌土曲霉转化
DOI：
10.1016/j.mimet.2014.01.007
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发表时间：
2014-03-01
影响因子：
2.2
通讯作者：
Wang, Li
中科院分区：
文献类型：
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作者：
Wang, Dongyang;He, Dan;Wang, Li
农杆菌介导转化（Agrobacterium tumefacians -mediated transformation， ATMT）已广泛应用于多种生物。在本研究中，我们开发了一种简单有效的利用ATMT对丝状真菌地曲霉进行遗传转化的系统。对转化方案进行了一定的参数优化，以快速生成田螺T-DNA插入突变体文库。通过PCR证实了有丝分裂稳定的潮霉素抗性基因（hph）在基因组中的整合，并利用热不对称交错PCR克隆了T-DNA侧翼序列。突变体文库的成功构建证明了ATMT方法对这种重要真菌的未来正向和反向遗传研究的实用性。(C) 2014 Elsevier B.V.版权所有
Agrobacterium tumefaciens-mediated transformation (ATMT) has been widely used in various organisms. In the current study, we developed a simple and efficient system for genetic transformation of the filamentous fungus Aspergillus terreus using ATMT. The transformation protocol was optimized for certain parameters to rapidly generate a library of Transferred DNA (T-DNA) insertion mutants of A. terreus. The presence of mitotically stable hygromycin resistance gene (hph) integration in the genome was confirmed by PCR, and T-DNA flanking sequences were cloned by thermal asymmetric interlaced PCR. The successful construction of the mutant library demonstrated the utility of the ATMT approach for future forward and reverse genetic studies in this important fungus. (C) 2014 Elsevier B.V. All rights reserved.