A novel improved method for Aspergillus nidulans transformation
A novel improved method for Aspergillus nidulans transformation
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DOI：
10.1016/s0167-7012(03)00208-2
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发表时间：
2003-12-01
影响因子：
2.2
通讯作者：
Diallinas, G
中科院分区：
文献类型：
--
作者：
Koukaki, M;Giannoutsou, E;Diallinas, G
我们系统地研究了利用从分生孢子萌发的不同阶段和幼菌丝体制备的原生质体转化构巢曲霉的效率。使用标准的整合质粒，获得增加的转化产量与原生质体分离从一个特定的阶段与芽管出现一致。根据使用的不同质粒，这种增加平均为2至8倍。用复制型质粒的转化效率与使用先前描述的方法获得的转化效率相似。尽管这一观察结果表明，转化效率的提高可能是由于质粒和基因组序列之间重组效率的提高，但我们不能排除与所使用的特定发育阶段相关的其他因素。在这项研究的过程中，我们还研究了可能提高转化率的其他参数的影响。所描述的方法也明显比其他现有方法更容易和更快。(C)2003 Elsevier B.V.保留所有权利。
We systematically investigated the efficiency of Aspergillus nidulans transformation using protoplasts prepared from different stages of conidiospore germination and young mycelium. Using standard integrative plasmids, increased transformation yields were obtained with protoplasts isolated from a specific stage coincident with germ tube emergence. This increase ranged, on the average, from two- to eightfold depending on different plasmids used. Transformation efficiencies with a replicative plasmid were similar to those obtained using previously described methods. Although this observation suggests that elevated transformation efficiencies might be due to increased efficiency of recombination between plasmid and genomic sequences, we cannot exclude other factors associated with the particular developmental stage used. In the course of this study, we also examined the effect of other parameters that might enhance transformation yields. The method described is also significantly easier and faster than other current methods. (C) 2003 Elsevier B.V. All rights reserved.