RNA silencing as a tool for exploring gene function in ascomycete fungi
RNA silencing as a tool for exploring gene function in ascomycete fungi
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DOI：
10.1016/j.fgb.2005.01.002
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发表时间：
2005-04-01
影响因子：
3
通讯作者：
Mayama, S
中科院分区：
文献类型：
--
作者：
Nakayashiki, H;Hanada, S;Mayama, S
我们开发了一种用于子囊菌真菌的 pHANNIBAL 样沉默载体 pSilent-1，它携带潮霉素抗性盒和用于发夹 RNA 表达的转录单位，并带有来自稻瘟病菌 Magnaporthe oryzae 的角质酶基因内含子的间隔区。在米霉中，具有角质酶内含子间隔区（147 bp）的沉默载体在沉默 eGFP 基因方面表现出比具有 GUS 基因片段间隔区或几丁质结合蛋白基因的较长内含子（850 bp）的沉默载体更高的效率。将pSilent-1应用于米霉的两个内源基因MPG1和聚酮合酶样基因，导致70-90%的所得转化体中的基因发生不同程度的沉默。基于 pSilent-1 的载体也在炭疽菌中诱导 RNA 沉默，其效率略低于米霉，表明该沉默载体应在子囊菌真菌中提供有用的反向遗传工具。 (c) 2005 Elsevier Inc. 保留所有权利。
We have developed a pHANNIBAL-like silencing vector, pSilent-1, for ascomycete fungi, which carries a hygromycin resistance cassette and a transcriptional unit for hairpin RNA expression with a spacer of a cutinase gene intron from the rice blast fungus Magnaporthe oryzae. In M. oryzae, a silencing vector with the cutinase intron spacer (147 bp) showed a higher efficiency in silencing of the eGFP gene than did those with a spacer of a GUS gene fragment or a longer intron (850 bp) of a chitin binding protein gene. Application of pSilent-1 to two M. oryzae endogenous genes, MPG1 and polyketide synthase-like gene, resulted in various degrees of silencing of the genes in 70-90% of the resulting transformants. RNA silencing was also induced by a pSilent-1-based vector in Colletotrichum lagenarium at a slightly lower efficiency than in M. oryzae, indicating that this silencing vector should provide a useful reverse genetic tool in ascomycete fungi. (c) 2005 Elsevier Inc. All rights reserved.