Agrobacterium-mediated transformation as a tool for functional genomics in fungi
Agrobacterium-mediated transformation as a tool for functional genomics in fungi
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DOI：
10.1007/s00294-005-0578-0
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发表时间：
2005-07-01
期刊：
影响因子：
2.5
通讯作者：
Ram, AFJ
中科院分区：
文献类型：
--
作者：
Michielse, CB;Hooykaas, PJJ;Ram, AFJ
在功能基因组学时代，对进行大规模靶向和随机诱变的工具的需求正在增加。一个潜在的工具是农杆菌介导的真菌转化。A.根癌农杆菌能够将其DNA的一部分（转移的DNA; T-DNA）转移到多种真菌中，并且可以通过农杆菌介导的转化（AMT）转化的真菌的数量仍在增加。AMT特别为难以用传统方法转化或传统方案未能产生稳定DNA整合的真菌开辟了分子遗传学领域。由于通过A.根癌农杆菌中，相对容易产生大量稳定的转化体。结合T-DNA随机整合并主要作为单拷贝的发现，AMT非常适合在真菌中进行插入诱变。此外，在各种基因打靶实验中，获得了较高的同源重组频率，表明T-DNA也是一种有用的靶向诱变底物。在这篇综述中，我们讨论了潜在的农杆菌DNA转移系统被用作一种工具，在真菌中的靶向和随机诱变。
In the era of functional genomics, the need for tools to perform large-scale targeted and random mutagenesis is increasing. A potential tool is Agrobacterium-mediated fungal transformation. A. tumefaciens is able to transfer a part of its DNA (transferred DNA; T-DNA) to a wide variety of fungi and the number of fungi that can be transformed by Agrobacterium-mediated transformation (AMT) is still increasing. AMT has especially opened the field of molecular genetics for fungi that were difficult to transform with traditional methods or for which the traditional protocols failed to yield stable DNA integration. Because of the simplicity and efficiency of transformation via A. tumefaciens, it is relatively easy to generate a large number of stable transformants. In combination with the finding that the T-DNA integrates randomly and predominantly as a single copy, AMT is well suited to perform insertional mutagenesis in fungi. In addition, in various gene-targeting experiments, high homologous recombination frequencies were obtained, indicating that the T-DNA is also a useful substrate for targeted mutagenesis. In this review, we discuss the potential of the Agrobacterium DNA transfer system to be used as a tool for targeted and random mutagenesis in fungi.