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Construction of pDESTR, a GATEWAY Vector for Gene Disruption in Filamentous Fungi
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)
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Submitter: 曾根 輝雄
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