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Construction of pDESTR, a GATEWAY Vector for Gene Disruption in Filamentous Fungi

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Title: Construction of pDESTR, a GATEWAY Vector for Gene Disruption in Filamentous Fungi
Authors: Abe, Ayumi Browse this author
Elegado, Evelyn B. Browse this author
Sone, Teruo3 Browse this author →KAKEN DB
Authors(alt): 曾根, 輝雄3
Issue Date: Mar-2006
Publisher: Springer
Journal Title: Current Microbiology
Volume: 52
Issue: 3
Start Page: 210
End Page: 215
Publisher DOI: 10.1007/s00284-005-0238-0
PMID: 16479358
Abstract: We have constructed pDESTR, a destination vector of gateway system especially for gene targeting and disruption in filamentous fungi. The vector was constructed by removing the multicloning site of pGEM-T easy vector, and inserting hygromycin phosphotransferase gene construct from pCB1004, and a gateway vector conversion cassette. In order to construct a DNA for gene disruption, only an inverse-polymerase chain reaction (PCR) amplification of the restricted, target sequence is needed. After the amplification with a 5′CACC-tagged primer and an ordinary primer, the DNA fragment will be inserted into pENTR/D-TOPO vector and then transferred into pDESTR through LR-recombination reaction. The resulting vector has the disruption construct, after being digested with the restriction enzyme used for the inverse-PCR. The effectiveness of this vector was assessed in Neurospora crassa. The use of pDESTR will therefore simplify the construction of a targeting vector, where multiple ligation steps are usually needed
Rights: The original publication is available at www.springerlink.com
Type: article (author version)
URI: http://hdl.handle.net/2115/5851
Appears in Collections:農学院・農学研究院 (Graduate School of Agriculture / Faculty of Agriculture) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 曾根 輝雄

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