Use of a nonhomologous end joining deficient strain (Δku70) of the ergot fungus Claviceps purpurea for identification of a nonribosomal peptide synthetase gene involved in ergotamine biosynthesis
Use of a nonhomologous end joining deficient strain (Δku70) of the ergot fungus Claviceps purpurea for identification of a nonribosomal peptide synthetase gene involved in ergotamine biosynthesis
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DOI：
10.1016/j.fgb.2007.04.008
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发表时间：
2008-01-01
影响因子：
3
通讯作者：
Tudzynski, Paul
中科院分区：
文献类型：
--
作者：
Haarmann, Thomas;Lorenz, Nicole;Tudzynski, Paul
麦角菌麦角菌麦角菌为了提高基因打靶效率，我们在两种不同的菌株中缺失了C purpurea ku 70基因：致病菌株20.1和非致病的麦角生物碱产生菌株P1。突变体的营养和病原体的发展，也没有生物碱生产受损。在Delta ku 70突变体中基因靶向效率显著增加（50-60%）。使用PI Delta ku 70菌株（生产麦角胺和麦角隐亭）靶向缺失lpsA 1，lpsA 1是生物碱基因簇中存在的两个三模块NRPS基因之一，编码参与麦角肽三肽部分形成的D-麦角酰肽合成酶。缺乏lpsA 1基因的突变体被证明不能产生麦角胺，但仍然能够产生麦角隐亭，证明LpsA 1参与麦角胺的生物合成。(C)2007爱思唯尔公司All rights reserved.
The ergot fungus Claviceps purpurea uses mainly the nonhomologous-end-joining (NHEJ) system for integration of exogenous DNA, leading to a low frequency of homologous integration (1-2%). To improve gene targeting efficiency we deleted the C purpurea ku70 gene in two different strains: the pathogenic strain 20.1 and the apathogenic, ergot alkaloid producing strain P1. The mutants were not impaired in vegetative and pathogenic development nor alkaloid production. Gene targeting efficiency was significantly increased (50-60%) in the Delta ku70 mutants. The PI Delta ku70 strain (producing ergotamine and ergocryptine) was used for targeted deletion of lpsA1, one of the two trimodular NRPS genes present in the alkaloid gene cluster, encoding D-lysergyl peptide synthetases involved in formation of the tripeptide moiety of ergopeptines. Mutants lacking the lpsA1 gene were shown to be incapable of producing ergotamine but were still able to produce ergocryptine, proving that LpsA1 is involved in ergotamine biosynthesis. (C) 2007 Elsevier Inc. All rights reserved.