An efficient system for heterologous expression of secondary metabolite genes in Aspergillus nidulans.
An efficient system for heterologous expression of secondary metabolite genes in Aspergillus nidulans.
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DOI：
10.1021/ja401945a
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发表时间：
2013-05-22
影响因子：
15
通讯作者：
Oakley, Bed R.
中科院分区：
文献类型：
--
作者：
Chiang, Yi-Ming;Oakley, C. Elizabeth;Ahuja, Manmeet;Entwistle, Ruth;Schultz, Aric;Chang, Shu-Lin;Sung, Calvin T.;Wang, Clay C. C.;Oakley, Bed R.
真菌次级代谢产物（SM）是具有医学价值的化合物的重要来源。基因组计划揭示，真菌有许多SM生物合成基因簇，通常不表达。为了获得这些潜在的有价值的，隐藏的集群，我们已经开发了一个异源表达系统在构巢曲霉。我们已经开发了一个有效的系统，用于从靶真菌扩增基因，将它们置于可调控的启动子的控制下，将它们转移到A。nidulans和表达它们。我们已经通过表达土曲霉的非还原聚酮酶和化合物生产和释放所需的其他基因来验证该系统。我们已经从这些NR-PKS中的六个获得了化合物的产生和释放，并鉴定了产物。为了证明该过程允许整个次级代谢物生物合成途径的转移和表达，我们表达了沉默A的所有基因。terreus集群并证明它产生Asperfuranone。此外，通过以各种组合表达该途径的基因，我们已经阐明了曲霉呋喃酮生物合成途径。我们还开发了删除整个A的程序。nidulans SM簇。这使我们能够去除可能干扰异源表达基因分析的簇，并消除不需要的毒素。
Fungal secondary metabolites (SMs) are an important source of medically valuable compounds. Genome projects have revealed that fungi have many SM biosynthetic gene clusters that are not normally expressed. To access these potentially valuable, cryptic clusters, we have developed a heterologous expression system in Aspergillus nidulans. We have developed an efficient system for amplifying genes from a target fungus, placing them under control of a regulatable promoter, transferring them into A. nidulans and expressing them. We have validated this system by expressing non-reducing polyketide synthases of Aspergillus terreus and additional genes required for compound production and release. We have obtained compound production and release from six of these NR-PKSs and have identified the products. To demonstrate that the procedure allows transfer and expression of entire secondary metabolite biosynthetic pathways, we have expressed all the genes of a silent A. terreus cluster and demonstrate that it produces asperfuranone. Further, by expressing the genes of this pathway in various combinations, we have clarified the asperfuranone biosynthetic pathway. We have also developed procedures for deleting entire A. nidulans SM clusters. This allows us to remove clusters that might interfere with analyses of heterologously expressed genes and to eliminate unwanted toxins.
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DOI：
10.1007/bf00339600
发表时间：
1988-08-01
期刊：
MOLECULAR AND GENERAL GENETICS
影响因子：
--
作者：
DUNNE, PW;OAKLEY, BR
通讯作者：
OAKLEY, BR
影响因子：
3.5
作者：
GWYNNE, DI;BUXTON, FP;SEALYLEWIS, HM
通讯作者：
SEALYLEWIS, HM
影响因子：
4.4
作者：
Chiang, Yi-Ming;Szewczyk, Edyta;Oakley, Berl R.
通讯作者：
Oakley, Berl R.
DOI：
10.1021/ci0200467
发表时间：
2003-01-01
期刊：
JOURNAL OF CHEMICAL INFORMATION AND COMPUTER SCIENCES
影响因子：
--
作者：
Feher, M;Schmidt, JM
通讯作者：
Schmidt, JM
影响因子：
14.8
作者：
Bok, Jin Woo;Chiang, Yi-Ming;Szewczyk, Edyta;Reyes-Domingez, Yazmid;Davidson, Ashley D.;Sanchez, James F.;Lo, Hsien-Chun;Watanabe, Kenji;Strauss, Joseph;Oakley, Berl R.;Wang, Clay C. C.;Keller, Nancy P.
通讯作者：
Keller, Nancy P.