Transformation of taxol-producing endophytic fungi by restriction enzyme-mediated integration (REMI).
The REMI method was used to introduce the plasmid pV2 harboring the hygromycin B phosphotransferase (hph) gene controlled by the Aspergillus nidulans trpC promoter and the trpC terminator into a taxol-producing endophytic fungus BT2. REMI transformation yielded stable transformants capable of continuing to grow on PDA medium containing 125 mug mL(-1) hygromycin B. The transformation efficiency was about 5-6 transformants mug(-1) plasmid DNA. The presence of hph gene in transformants was confirmed by PCR and Southern blot analyses. To the authors' knowledge, this is the first report on the transformation of taxol-producing endophytic fungi by the REMI technique. This study provides an effective approach for improving taxol production of endophytic fungi by the genetic engineering of taxol biosynthetic pathway genes in the future.
Wang
Yechun
Y
Plant Biotechnology Research Center, School of Agriculture and Biology, School of Life Science and Technology, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Shanghai Jiao Tong University, Shanghai, China.
Guo
Binhui
B
Miao
Zhiqi
Z
Tang
Kexuan
K
D000935:Antifungal Agents / Q000494:pharmacology; D001203:Ascomycota / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D001233:Aspergillus nidulans / Q000235:genetics; D015139:Blotting, Southern; D025141:Drug Resistance, Fungal / Q000235:genetics; D018014:Gene Transfer Techniques; D005818:Genetic Engineering; D006921:Hygromycin B / Q000494:pharmacology; D017239:Paclitaxel / Q000096:biosynthesis*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D014170:Transformation, Genetic
D000935:Antifungal Agents; D006921:Hygromycin B; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase; D017239:Paclitaxel