[大丽轮枝菌中的高效基因敲除]
[High efficient gene knockout in Verticillium dahliae].
作者信息
Tian Li, Chen Jieyin, Wang Jiani, Wang Jinlong, Dai Xiaofeng
机构信息
Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
出版信息
Wei Sheng Wu Xue Bao. 2011 Jul;51(7):906-13.
OBJECTIVE
We developed an efficient method of gene knockout in Verticillium dahliae, an important soil-borne fungal pathogen that causes cotton vascular wilt diseases.
METHODS
By using fusion PCR, we constructed gene knockout vectors. By using Agrobacterium tumefaciens-mediated transformation and applying a herpes simplex virus thymidine kinase (HSVtk) gene in T-DNA as a conditional lethal gene to counter-select against ectopic transformants, we developed an efficient method to select gene knockout transformants.
RESULTS
Gene knockout frequency for ADE4 and ChsV was 87% and 44% , respectively.
CONCLUSION
We developed an efficient tool for gene knockout in Verticillium dahliae, which would help clarify the infection mechanism of this fungal pathogen.
目的
我们开发了一种在大丽轮枝菌中进行基因敲除的有效方法，大丽轮枝菌是一种重要的土传真菌病原体，可导致棉花维管束枯萎病。
方法
通过融合PCR构建基因敲除载体。利用根癌农杆菌介导的转化，并在T-DNA中应用单纯疱疹病毒胸苷激酶（HSVtk）基因作为条件致死基因来反选异位转化体，我们开发了一种选择基因敲除转化体的有效方法。
结果
ADE4和ChsV的基因敲除频率分别为87%和44%。
结论
我们开发了一种在大丽轮枝菌中进行基因敲除的有效工具，这将有助于阐明这种真菌病原体的感染机制。
Zotero 插件