title: Agrobacterium tumefaciens-mediated transformation of a taxol-producing endophytic fungus, Cladosporium cladosporioides MD2.
abstract: In this study, an Agrobacteriurn tumefaciens-mediated transformation (ATMT) protocol was successfully developed for the genetic transformation of a taxol-producing fungus, Cladosporium cladosporioides MD2, and the co-cultivation conditions affecting the transformation efficiency were optimized. The optimal transformation conditions were that 1 ml of C. cladosporioides MD2 spore suspension (10(8) spores/ml) was mixed with an equal volume of A. tumefaciens cultures, which contained 400 μl of A. tumefaciens LBA4404 (OD(660) ≈ 0.6) and 600 μl LB medium that were used to make up difference in volume, and the mix cultures were supplemented with 300 μM acetosyringone (AS) and co-cultivated at 26°C and 50 rpm for 48 h. Stable transformants were obtained through analysis of the mitotic stability of inserted T-DNA and the presence of hygromycin resistance gene (hpt II). This study laid a fine groundwork for development of transgenic C. cladosporioides MD2 strains.
authors.affiliation: Institute of Resource Biology and Biotechnology, Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Zhang
authors.forename: Ting-Ting
authors.initials: TT
authors.lastname: Liu
authors.forename: Peng-Peng
authors.initials: PP
authors.lastname: Zhou
authors.forename: Shu-Tao
authors.initials: ST
authors.lastname: Li
authors.forename: Long-Jiang
authors.initials: LJ
authors.lastname: Yu
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics*; D002963:Cladosporium / Q000235:genetics* / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D017239:Paclitaxel / Q000378:metabolism*; D014170:Transformation, Genetic
chemical_list: D004269:DNA, Bacterial; C036483:T-DNA; D017239:Paclitaxel